ES2724801T3 - Combinations of anti-4-1BB antibodies and ADCC inducing antibodies for cancer treatment - Google Patents

Abstract

An anti-4-1BB antibody, or an antigen binding portion thereof, for use in the treatment of cancer in combination with an anti-CD20 antibody, or an antigen binding portion thereof, wherein said anti- antibody 4-1BB, or the antigen binding portion thereof, comprises: (a) an H-CDR1 as set forth in SEQ ID NO: 29; (b) an H-CDR2 as set forth in SEQ ID NO: 30; (c) an H-CDR3 as set forth in SEQ ID NO: 31; (d) an L-CDR1 as set forth in SEQ ID NO: 34; (e) an L-CDR2 as set forth in SEQ ID NO: 35; and (f) an L-CDR3 as set forth in SEQ ID NO: 36, and wherein the anti-CD20 antibody, or antigen binding portion thereof, is administered in a dose of 3-15 mg / kg. .

Description

DESCRIPTION

Combinations of anti-4-1BB antibodies and ADCC inducing antibodies for cancer treatment Background

Cancer is now the leading cause of death in the United States. Currently, it is usually treated with one or a combination of three types of therapies: surgery, radiation and chemotherapy. Chemotherapy involves disruption of cellular replication or cellular metabolism. The adverse effects of systemic chemotherapy used in the treatment of neoplastic disease can be life-threatening and have become of great importance for the clinical treatment of cancer patients.

4-1BB (also called CD137, TNFRSF9, etc.) is a transmembrane protein of the tumor necrosis factor receptor superfamily (TNFRS). The current understanding of 4-1BB indicates that expression generally depends on activation and is present in a broad subset of immune cells that include activated NK and NKT cells, regulatory T cells, dendritic cells (DC), stimulated mast cells, differentiating myeloid cells , monocytes, neutrophils and eosinophils (Wang, 2009, Immunological Reviews 229: 192-215). The expression of 4-1BB has also been demonstrated in the tumor vasculature (Broil, 2001, Amer. J Clin. Pathol. 115 (4): 543-549; Seaman, 2007, Cancer Cell 11: 539-554) and in atherosclerotic sites of inflammation or endothelium (Drenkard, 2007 FASEB J. 21: 456-463; Olofsson, 2008, Circulation 117: 1292-1301). The ligand that stimulates 4-1BB, that is, 4-1BB ligand (4-1BBL), is expressed in activated antigen presenting cells (APC), myeloid progenitor cells and hematopoietic stem cells.

Human 4-1BB is a 255 amino acid protein (accession number NM_001561; NP_001552). The complete human 4-1BB amino acid sequence is provided in SEQ ID NO: 68. The protein comprises a signal sequence (amino acid residues 1-17), followed by an extracellular domain (169 amino acids), a transmembrane region (27 amino acids) and an intracellular domain (42 amino acids) (Cheuk ATC et al. 2004 Cancer Gene Therapy 11: 215-226). The receptor is expressed on the cell surface in the form of a monomer and dimer and is probably trimerized with a 4-1BB ligand for signaling.

Numerous studies of murine and human T cells indicate that 4-1BB promotes increased cell proliferation, survival and cytokine production (Croft, 2009, Nat Rev Immunol 9: 271-285). Studies have indicated that some 4-1BB agonist mAbs increase the expression of the costimulatory molecule and significantly improve the responses of cytolytic T lymphocytes, resulting in antitumor efficacy in several models. 4-1BB agonist mAbs have demonstrated efficacy in prophylactic and therapeutic environments. In addition, models of monotherapy tumors with 4-1BB and combination therapy have established memory responses of durable antitumor protective T cells (Lynch, 2008, Immunol Rev. 22: 277-286). 4-1BB agonists have also been shown to inhibit autoimmune reactions in a variety of autoimmunity models recognized in the art (Vinay, 2006, J Mol Med 84: 726-736). This dual 4-1BB activity offers the potential to provide antitumor activity while reducing autoimmune side effects that may be associated with immunotherapy approaches that break immune tolerance.

The development of targeted therapies focuses on the specific orientation of neoplastic cells while preserving normal tissues to lessen side effects. An alternative and / or additional approach to cancer therapy is to target the immune system instead of and / or in addition to targeting the tumor itself. It has been hypothesized that ADCC is a mechanism of tumor destruction that results in the direct presentation of the antigen and in the induction of specific T-cell responses of the tumor antigen ("cross-priming") (Weiner et al., 2009 ).

There is a long unmet need for antibodies that bind to human 4-1BB, increasing the response mediated by 4-1BB and, therefore, provide therapeutic potential for the treatment of various diseases and conditions, including cancer.

Kohrt et al., Blood, 2001 117: 2423-2432 describes the use of an anti-CD137 antibody (anti-4-1BB) in combination with an anti-CD20 antibody that indicates anti-lymphoma activity in vivo.

Summary

The present invention provides an anti-4-1BB antibody, or an antigen binding portion thereof, for use in the treatment of cancer in combination with an anti-CD20 antibody, or an antigen binding portion thereof, in the that said anti-4-1BB antibody, or antigen binding portion thereof, comprises:

(a) an H-CDR1 as set forth in SEQ ID NO: 29;

(b) an H-CDR2 as set forth in SEQ ID NO: 30;

(c) an H-CDR3 as set forth in SEQ ID NO: 31;

(d) an L-CDR1 as set forth in SEQ ID NO: 34;

(e) an L-CDR2 as set forth in SEQ ID NO: 35; Y

(f) an L-CDR3 as set forth in SEQ ID NO: 36;

and wherein the anti-CD20 antibody, or its antigen binding portion, is administered in a dose of approximately 3-15 mg / kg.

In some embodiments, the anti-CD20 antibody, or its antigen binding portion, comprises the 6 CDRs of rituximab. In other embodiments, the anti-4-1BB antibody, or the antigen-binding portion thereof, comprises a Vh region comprising the amino acid sequence set forth in SEQ ID NO: 43 and a Vl region comprising the amino acid sequence set forth in SEQ ID NO: 45. In further embodiments, the anti-CD20 antibody, or its antigen binding portion, comprises the V ^h region of rituximab and the V ^l region of rituximab. In further embodiments, the anti-4-1BB antibody, or the antigen binding portion thereof, comprises a heavy chain amino acid sequence as set forth in SEQ ID NO: 44 and further comprises a light chain amino acid sequence. as set forth in SEQ ID NO: 46, with the proviso that the C-terminal end lysine residue of SEQ ID NO: 44 is optionally absent. In some embodiments, the anti-CD20 antibody, or its antigen binding portion, comprises the rituximab heavy chain amino acid sequence and the rituximab light chain amino acid sequence. In more embodiments, the method comprises administering to said patient a therapeutically effective amount of MOR-7480.1, or an antigen binding portion thereof, in combination with a therapeutically effective amount of rituximab, or an antigen binding portion thereof.

Brief description of the drawings

Figure 1 shows the combinatorial efficacy of monoclonal antibodies 4-1BB and CD20 in a lymphoma model.

Figure 2 shows the combinatorial efficacy of a 4-1BB monoclonal antibody and a P-cadherin monoclonal antibody (g-194-g09 or anti-P-cadherin1) in a colon carcinoma model.

Figure 3 shows the combinatorial efficacy of a 4-1BB monoclonal antibody and a P-cadherin (anti-P-cadherin2) monoclonal antibody in a colon carcinoma model.

Figures 4A and 4B illustrate that combination therapy with monoclonal antibodies 4-1BB and CD20 reduces the circulating tumor burden in a spontaneous murine model of lymphoma.

Figure 5 shows the survival benefit of the combinatorial treatment of mice with jmyc E lymphoma with monoclonal antibodies 4-1BB and CD20.

Detailed description

The embodiments described herein refer to the treatment of cancer in a patient in need of such treatment using anti-4-1BB antibodies (eg, MOR-7480.1) in combination with one or more antibodies that induce antibody-dependent cellular cytotoxicity. (ADCC) (for example, an anti-CD20 antibody such as rituximab). Those skilled in the art will recognize that certain subclasses of antibodies, such as IgG1 and IgG3, induce ADCC activity. ADCc activity may also be the result of the introduction of carbohydrate modifications, such as altered glycosylation patterns, in the Fc region compared to the pattern of native carbohydrates. Those skilled in the art also recognize that certain mutations in the Fc region of an antibody, compared to the wild-type Fc region, increase ADCC activity. The present disclosure demonstrates the combinatorial efficacy of anti-4-1BB antibodies and ADCC inducing antibodies in tumor models.

Unless otherwise defined herein, the scientific and technical terms used in connection with the present embodiments will have the meanings commonly understood by those skilled in the art. In addition, unless the context requires otherwise, the singular terms will include the plurals and the plural terms will include the singular. In general, the nomenclatures used in relation to, and the techniques of cell and tissue culture, molecular biology, immunology, microbiology, genetics and chemistry of nucleic acids and proteins and hybridization described herein are those well known and commonly used in the technique.

The procedures and techniques of the present embodiments are generally performed in accordance with procedures well known in the art and as described in several general and more specific references cited and discussed throughout the present specification, unless indicated otherwise. These references include, for example, Sambrook and Russell, Molecular Cloning, A Laboratory Approach, Cold Spring Harbor Press, Cold Spring Harbor, NY (2001), Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, NY ( 2002), and Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1990). Enzymatic reactions and purification techniques are performed in accordance with the manufacturer's specifications, as commonly performed in the art or as described herein. The nomenclatures used in relation to the laboratory procedures and techniques of analytical chemistry, synthetic organic chemistry and pharmaceutical and medicinal chemistry described herein are those well known and commonly used in the art. Standard techniques are used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and administration, and treatment of patients.

Definitions

As used herein, each of the following terms has the meaning associated in this section.

The articles "a" and "one, one" are used herein to refer to one or more than one (ie, at least one) of the grammatical object of the article. As an example, "an element" means an element or more than one element.

As used herein, the twenty conventional amino acids and their abbreviations follow the conventional use. See Immunology - A Synthesis (2nd edition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)).

A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain R group with similar chemical properties (eg, loading or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percentage of sequence identity or the degree of similarity can be adjusted up to correct the conservative nature of the substitution. The means for making this adjustment are well known to those skilled in the art. See, for example, Pearson, Methods Mol. Biol. 243: 307-31 (1994). Examples of amino acid groups having side chains with similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) side chains containing amide: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine and tryptophan; 5) basic side chains: lysine, arginine and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine and methionine. Preferred conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalaninatyrosine, lysine-arginine, alanine-valine, glutamate-aspartate and asparagine-glutamine.

Alternatively, a conservative substitution is any change that has a positive value in the logarithmic probability matrix PAM250 disclosed in Gonnet et al., Science 256: 1443-45 (1992). A "moderately conservative" replacement is any change that has a non-negative value in the logarithmic probability matrix PAM250.

Preferred amino acid substitutions are those that: (1) reduce the susceptibility to proteolysis, (2) reduce the susceptibility to oxidation, (3) alter the binding affinity to form protein complexes, and (4) confer or modify other physicochemical or functional properties of such analogs. Analogs comprising substitutions, deletions and / or insertions may include several muteins of a sequence other than the natural peptide sequence. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) can be made in the sequence that occurs naturally (preferably in the portion of the polypeptide outside the domain or domains that form intermolecular contacts). A conservative amino acid substitution should not substantially change the structural characteristics of the main sequence (for example, a replacement amino acid should not tend to break a helix that occurs in the main sequence, or interrupt other types of secondary structure that characterize the sequence principal). Examples of secondary and tertiary polypeptide structures recognized in the art are described in Proteins, Structures and Molecular Principles (Creighton, Ed., W. H. Freeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et al., Nature 354: 105 (1991).

Sequence similarity for polypeptides, which is also called sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measures assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For example, GCG contains programs such as "Gap" and "Bestfit" that can be used with predetermined parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides of different species of organisms or between a protein wild type and a mutein thereof. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA using predetermined or recommended parameters, a program in GCG version 6.1. FASTA (for example, FASTA2 and FASTA3) provides alignments and sequence identity percentages of the regions of the best overlap between the question and search sequences (Pearson, Methods Enzymol. 183: 63-98 (1990); Pearson, Methods Mol Biol. 132: 185-219 (2000)). Another preferred algorithm when comparing a sequence of the invention with a database containing a large number of sequences from different organisms is the BLAST software, especially blastp or tblastn, using predetermined parameters. See, for example, Altschul et al., J. Mol. Biol. 215: 403-410 (1990); Altschul et al., Nucleic Acids Res. 25: 3389-402 (1997).

An intact "antibody" comprises at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. See generally, Fundamental Immunology, chapter 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)). Each heavy chain is composed of a heavy chain variable region (HCVR or Vh) and a heavy chain constant region (Ch). The constant region of the heavy chain is composed of three domains, CH1, CH2 and CH3. Each light chain is composed of a variable light chain region (LCVR or V ^l ) and a constant light chain region. The constant region of the light chain is composed of a domain, C ^l . Regions V ^h and Vl can be subdivided into regions of hypervariability, called complementarity determining regions (CDR), interspersed with regions that are more conserved, called framework regions (FR). Each V ^h and V ^l are composed of three CDRs and four FRs, arranged from the amino terminal end to the carboxyl terminal end in the following order: FR1, CDR1, ^f R2, CDR2, FR3, CDR3, FR4. The assignment of amino acids to each domain is in accordance with the definitions of Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD (1987 and 1991)), or Chothia & Lesk, J. Mol. Biol. 196: 901-917 (1987); Chothia et al., Nature 342: 878-883 (1989).

The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. Constant regions of the antibodies can mediate the binding of immunoglobulin to tissues or host factors, including several cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.

The term "antibody" may include antigen-binding portions of an intact antibody that retain ability to specifically bind to the intact antibody antigen (eg, 4-1BB, CD20 or P-cadherin). Antigen binding portions can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.

Examples of antigen binding portions include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) an F (ab ') 2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge in the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; (iv) an Fv fragment consisting of the single-arm VL and VH domains of an antibody, (v) a single domain antibody ("dAb"), which consists of a VH domain as described in Ward et al. ., Nature 341: 544-546 (1989); and (vi) an isolated complementarity determining region (CDR). In addition, although the two domains of the Fv fragment, V ^h and V ^l , are encoded by separate genes, they can be linked, by recombinant procedures, by a synthetic linker that allows them to be manufactured as a single protein chain in which the V regions ^h and V ^l mate to form monovalent molecules (known as single chain Fv (scFv); see, for example, Bird et al., Science 242: 423-426 (1988); and Huston et al., Proc. Natl Acad. Sci. USA 85: 5879-5883 (1988)). Such single chain antibodies are included by reference to the term "antibody."

A "bispecific antibody" has two different binding specificities, see, for example, United States Patent No. 5,922,845 and United States Patent No. 5,837,243; Zeilder J. Immunol. 163: 1246-1252 (1999); Somasundaram Hum. Antibodies 9: 47-54 (1999); Keler Cancer Res. 57: 4008-4014 (1997). For example, the invention provides bispecific antibodies that have a binding site for a cell surface antigen (such as human 4-1BB, CD20 or P-cadherin), and a second binding site for an Fc receptor on the surface of An effector cell. The invention also provides multispecific antibodies, which have at least three binding sites.

The term "bispecific antibodies" also includes "diabodies." Diabodies are bivalent, bispecific antibodies, in which the V ^h and V ^l domains are expressed in a single polypeptide chain, but use a connector that is too short to allow pairing between the two domains in the same chain, forcing the domains to pair with complementary domains of another chain and create two antigen binding sites (see, for example, Holliger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993); Poljak et al. , Structure 2: 1121 1123 (1994)).

The terms "human antibody" or "human sequence antibody", as used interchangeably herein, include antibodies that have variable and constant regions (if present) derived from human germline immunoglobulin sequences. The human sequence antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (eg, mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody," as used herein, is not intended to include "chimeric" antibodies in which the CDR sequences derived from the germ line of another mammalian species, such as a mouse, have been grafted. in human framework sequences (ie "humanized" or PRIMATIZEDmr antibodies).

The term "chimeric antibody" as used herein means an antibody that comprises regions of two or more different antibodies. In one embodiment, one or more of the CDRs are derived from a human antibody. In another embodiment, all CDRs are derived from a human antibody. In another embodiment, the CDRs of more than one human antibody are combined in a chimeric human antibody. For example, an antibody chimeric may comprise a CDR1 of the light chain of a first human anti-4-1BB antibody, a CDR2 of the light chain of a second human anti-4-1BB antibody and a CDR3 and CDR3 of the light chain of a third human antibody anti-4-1BB, and heavy chain CDRs can be derived from one or more other anti-4-1BB antibodies. In addition, the framework regions may be derived from one of the same anti-4-1BB antibodies or from one or more different human or human.

In addition, as discussed hereinbefore, the chimeric antibody includes an antibody comprising a portion derived from the germline sequences of more than one species.

By the term "effective amount," or "therapeutically effective amount," as used herein, is meant an amount that when administered to a mammal, preferably a human being, mediates a detectable therapeutic response compared to the detected response. in the absence of the compound. A therapeutic response, such as, but not limited to, inhibition and / or decrease in tumor growth, tumor size, metastasis and the like, can easily be evaluated by a large number of procedures recognized in the art, including , for example, procedures such as those disclosed herein.

The person skilled in the art would understand that the effective amount of the compound or composition administered herein varies and can easily be determined based on a number of factors such as the disease or condition being treated, the stage of the disease, age and the health and physical condition of the mammal being treated, the severity of the disease, the particular compound being administered and the like.

A "therapeutically effective amount", or "effective amount" is intended to qualify the amount of an agent required to detectably reduce one or more of the symptoms of a neoplasia disorder, including, but not limited to: 1 ) reduction in the number of cancer cells; 2) reduction of tumor size; 3) inhibition (ie, deceleration to some extent, preferably arrest) of infiltration of cancer cells in peripheral organs; 3) inhibition (ie, deceleration to some extent, preferably arrest) of tumor metastasis; 4) inhibition, to some extent, of tumor growth; 5) relieve or reduce to some extent one or more of the symptoms associated with the disorder; and / or 6) alleviate or reduce the side effects associated with the administration of anticancer agents.

By the term "compete", as used herein with respect to an antibody, it is understood that a first antibody, or an antigen binding portion thereof, competes for binding with a second antibody, or a binding portion a antigen thereof, in which the binding of the first antibody with its related epitope is detectably reduced in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. The alternative, in which the binding of the second antibody to its epitope is also detectably reduced in the presence of the first antibody, may, but not necessarily, be the case. That is, a first antibody can inhibit the binding of a second antibody to its epitope without that second antibody inhibiting the binding of the first antibody to its respective epitope. However, when each antibody detectably inhibits the binding of the other antibody with its epitope or related ligand, whether in the same, greater or lesser extent, it is said that the antibodies "cross-compete" with each other for the binding of their epitope or respective epitopes. For example, cross-competing antibodies can bind to the epitope, or portion of the epitope, to which the antibodies of the embodiments bind. Both antibodies that compete and cross-compete are encompassed by the present embodiments. Regardless of the mechanism by which such competition or cross-competition occurs (for example, steric hindrance, conformational change or binding to a common epitope, or portion thereof, and the like), the person skilled in the art would appreciate, based on the teachings provided. herein, that such antibodies that compete and / or cross-compete are included and may be useful for the procedures described herein.

The term "epitope" includes any determinant protein capable of specifically binding to an immunoglobulin or T-cell receptor. Epitopic determinants generally consist of groups of chemically active molecules on the surface, such as amino acids or sugar side chains, and generally they have specific three-dimensional structural characteristics, as well as specific load characteristics. Conformational and non-conformational epitopes are distinguished because binding to the former, but not the latter, is lost in the presence of denaturing solvents.

The "instructional material", as said term is used herein, includes a publication, a record, a diagram, or any other means of expression that can be used to communicate the usefulness of the compound, combination and / or composition of the invention in the kit to affect, relieve or treat the various diseases or disorders mentioned herein. Optionally, or alternatively, the instructional material may describe one or more procedures to alleviate diseases or disorders in a cell, tissue or mammal, including those described elsewhere in this document.

The instructional material of the kit can, for example, be fixed to a container containing the compound and / or composition of the invention or can be sent together with a container containing the compound and / or composition.

Alternatively, the instructional material can be sent separately from the container with the intention that the recipient uses the instructional material and the compound cooperatively.

Except where indicated, the terms "patient" or "subject" are used interchangeably and refer to mammals such as human patients and non-human primates, as well as veterinary subjects such as rabbits, rats, mice and other animals. Preferably, patient refers to a human being.

The phrase "pharmaceutically acceptable salt or salts", as used herein, includes salts of acidic or basic groups that may be present in a compound. Compounds that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, that is, salts containing pharmacologically acceptable anions, such as acetate, benzenesulfonate, benzoate salts. , bicarbonate, bisulfate, bistosylate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dichlorohydrate, edetate, edisilate, stolate, esylate, ethyl succinate, fumarate, gluceptate, gluconate, glutacetate, glutacetate, glutamate, glycinate , hydrabamine, hydrobromide, hydrochloride, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsilate, nitrate, oleate, oxalate, pamoate, (embonate), palmitate, pantothenate / phosphate, phosphate , polygalacturonate, salicylate, stearate, subacetate, succinate, tanato, tartrate, teoclate, tosylate, thiodedode and valerate. Preferred salts of compounds 1-3 are disclosed in PCT Publication No. 2003/016305, United States Patent Application No. 10 / 956,420, filed September 30, 2004, and PCT Application No. PCT / IB2004 / 003070, filed September 20, 2004. Particularly preferred salts of compound 1 include malate salts, most preferably an L-malate salt. A particularly preferred salt of compound 3 is a maleate salt.

Conventional notation is used herein to represent polypeptide sequences: the left end of a polypeptide sequence is the amino terminal end; the right end of a polypeptide sequence is the carboxyl terminal end.

By the phrase "specifically binds", as used herein, is meant a compound, for example, a protein, a nucleic acid, an antibody and the like, which recognizes and binds to a specific molecule, but does not recognize substantially or binds to other molecules in a sample. For example, an antibody or a peptide inhibitor that recognizes and binds to a 4-1BB related ligand in a sample, but does not recognize or binds substantially to other molecules in the sample. Therefore, under the designated test conditions, the specified binding fraction (for example, an antibody or an antigen binding portion thereof) preferably binds to a particular target molecule and does not bind in a significant amount to others. components present in a test sample. A variety of assay formats can be used to select an antibody that specifically binds to a molecule of interest. For example, solid phase ELISA immunoassay, immunoprecipitation, BIAcore analysis and Western blotting are used to identify an antibody that reacts specifically with 4-1BB. Typically, a specific or selective reaction will be at least twice the signal or background noise and more typically more than 10 times the background, even more specifically, it is said that an antibody "specifically binds" to an antigen when the constant of equilibrium dissociation (KD) is <1 pM, preferably <100 nM and most preferably <10 nM.

The term "KD" refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

The term "antibody-dependent cell cytotoxicity" or "ADCC" refers to a cell-mediated reaction in which non-specific cytotoxic cells (eg, NK cells, neutrophils, macrophages, etc.) recognize the antibody bound to a cell. target and subsequently cause lysis of the target cell. Such cytotoxic cells that mediate ADCC generally express Fc receptors (FcR). Primary cells to mediate ADCC (NK cells) express F ^{cy and} RIII, while monocytes express F ^cy RI, F ^cy RII, F ^cy RIII and / or F ^cy RIV. To evaluate the ADCC activity of a molecule, an in vitro CCAD assay can be performed, such as that described in U.S. Patent Nos. 5,500,362 or 5,821,337. Effector cells useful for such assays include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK). Alternatively, or additionally, the ADCC activity of the molecules of interest can be evaluated in vivo , for example, in an animal model.

The term "ADCC inducing antibody" refers to an antibody that demonstrates ADCC as measured by assays known to those skilled in the art. Such activity is typically characterized by the union of the Fc region with several FcRs. Without being limited by any particular mechanism, those skilled in the art will recognize that the ability of an antibody to demonstrate ADCC can be, for example, by virtue of its subclass (such as IgG1 or IgG3), by mutations introduced into the Fc region, or under modifications to carbohydrate patterns in the Fc region of the antibody. Such modifications are described, for example, in United States Patent Publication No. 2007-0092521.

The antibody referred to herein as "rituximab" will be readily recognized by those skilled in the art, and is sold under the trade names Rituxan® and MabThera®. Rituximab is a genetically modified chimeric monoclonal antibody directed against the human CD20 antigen. This chimeric antibody contains a constant domain of human IgG1 and is identified by the name "C2B8" in US Patent No. 5,736,137 (Andersen, KC, et al.) Published April 17, 1998. Rituximab is approved for the treatment of patients with non-Hodgkin B-cell lymphoma, positive for CD20, relapsing or refractory of low or follicular grade. Studies of mechanisms of action in vitro have shown that rituximab shows human complement-dependent cytotoxicity (CDC), as well as significant activity in assays that measure antibody-dependent cellular cytotoxicity (ADCC).

As used herein, "substantially pure" means that a target species is the predominant species present (ie, on a molar basis it is more abundant than any other individual species in the composition), and preferably a substantially purified fraction is a composition. wherein the target species (for example, an anti-4-1BB antibody) comprises at least about 50 percent (on a molar basis) of all macromolecular species present. In general, a substantially pure composition will comprise more than about 80 percent of all macromolecular species present in the composition, more preferably more than about 85%, 90%, 95% and 99%. More preferably, the target species is purified to essential homogeneity (the contaminating species cannot be detected in the composition by conventional detection procedures), in which the composition consists essentially of a single macromolecular species.

As used herein, "treating" means reducing the frequency with which the symptoms of a disease (ie, tumor growth and / or metastasis, or other effect mediated by numbers and / or activity of the immune cells, and the like) are experienced by a patient. The term includes the administration of the compounds or agents of the present invention to prevent or delay the onset of symptoms, complications or biochemical signs of a disease (eg, elevated PSA level), relieve symptoms or stop or inhibit the further development of the disease, condition or disorder. The treatment can be prophylactic (to prevent or delay the onset of the disease, or to prevent the manifestation of clinical or subclinical symptoms thereof) or therapeutic suppression or relief of symptoms after the manifestation of the disease.

"Combination therapy" encompasses the administration of an ADCC-inducing antibody and a 4-1BB antibody as part of a specific treatment regimen intended to provide a beneficial effect of the joint action of these therapeutic agents. The beneficial effect of the combination includes, among others, the joint pharmacokinetic or pharmacodynamic action resulting from the combination of therapeutic agents. The administration of these therapeutic agents in combination is usually carried out for a defined period of time (generally minutes, hours, days or weeks, depending on the combination selected). "Combination therapy" is generally not intended to encompass the administration of two or more of these therapeutic agents as part of separate monotherapy regimens that result incidentally and arbitrarily in the combinations of the present invention. "Combination therapy" encompasses the administration of these therapeutic agents in a sequential manner, that is, in which each therapeutic agent is administered at a different time, as well as the administration of these therapeutic agents, or at least two of the agents. therapeutic, substantially simultaneously. Substantially simultaneous administration can be achieved, for example, by administering to the subject a single capsule having a fixed proportion of each therapeutic agent or in multiple individual capsules for each of the therapeutic agents. The sequential or substantially simultaneous administration of each therapeutic agent can be carried out by any appropriate route that includes, among others, oral routes, intravenous routes, intramuscular routes and direct absorption through the tissues of the mucous membrane. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the selected combination may be administered by intravenous injection, while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, both therapeutic agents can be administered orally or both therapeutic agents can be administered by intravenous injection. The sequence in which therapeutic agents are administered is not strictly critical. "Combination therapy" may also encompass the administration of therapeutic agents as described above in additional combination with other biologically active ingredients (such as, among others, a second and different antineoplastic agent) and non-pharmacological therapies (such as, but not limited to surgery or radiotherapy). When the combination therapy further comprises radiotherapy, the radiotherapy can be carried out at any suitable time provided that a beneficial effect of the joint action of the combination of the therapeutic agents and radiotherapy is achieved. For example, in appropriate cases, the beneficial effect is still achieved when radiotherapy is temporarily eliminated from the administration of therapeutic agents, perhaps for days or even weeks.

Description

The embodiments described herein refer to new therapeutic uses and medical procedures comprising the co-administration of a combination of an anti-4-1BB antibody and an ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab ) for the treatment of cancer in a patient who needs such treatment. In one embodiment, the method comprises administering an anti-4-1BB antibody (for example, MOR-7480.1) in combination with an anti-CD20 antibody (for example, rituximab).

I. Anti-4-1BB Antibodies

The preferred anti-4-1BB antibody is an antibody (for example, MOR-7480.1, MOR-7480, MOR-7480.2) that specifically binds to human 4-1BB and comprises:

(a) an H-CDR1 as set forth in SEQ ID NO: 29;

(b) an H-CDR2 as set forth in SEQ ID NO: 30;

(c) an H-CDR3 as set forth in SEQ ID NO: 31;

(d) an L-CDR1 as set forth in SEQ ID NO: 34;

(e) an L-CDR2 as set forth in SEQ ID NO: 35; Y

(f) an L-CDR3 as set forth in SEQ ID NO: 36.

Other antibodies of the disclosure include, but are not limited to, MOR-6032, MOR-7361, MOR-7483, MOR-7483.1 and MOR-7483.2. These anti-4-1BB antibodies are described in Table 1 and Table 2 below.

-

-

The amino acid and nucleic acid sequences of MOR-6032, MOR-7361, MOR-7480, MOR-7480.1, MOR-7480.2, MOR-7483, MOR-7483.1 and MOR-7483.2 are presented herein. Briefly, anti-4-1BB antibodies include antibodies that have amino acid sequences of the heavy and light chains of an antibody such as, for example, MOR-6032, MOR-7361, MOR-7480, MOR-7480.1, MOR-7480.2 , MOR-7483, MOR-7483.1 and MOR-7483.2. The embodiments described herein also refer to antibodies that have the amino acid sequences of the CDRs of the heavy and light chains of these antibodies. Other embodiments relate to anti-4-1BB antibodies that have the variable regions of the heavy and light chains of those antibodies. In another embodiment of the invention, the antibody is selected from an antibody having the full length variable region, CDR, amino acid sequences of the heavy and light chains of the MOR-7480.1, MOR-7480 or MOR-7480.2 antibody. Other antibodies of the disclosure have the full length variable region, or CDR, the heavy and light chains of the MOR-6032, MOR-7361, MOR-7483, MOR-7483.1 and MOR-7483.2 antibodies. In some embodiments, the anti-4-1BB antibody may comprise a or more of the sequences described above with one or more conservative substitutions. In another description, the anti-4-1BB antibody may be an antibody that cross-competes with MOR-6032, MOR-7361, MOR-7480, MOR-7480.1, MOR-7480.2, MOR-7483, MOR-7483.1, or MOR-7483.2.

In another embodiment, the methods described herein are practiced using an anti-4-1BB antibody comprising a heavy chain comprising the amino acid sequences of H-CDR1, H-CDR2 and H-CDR3, and a light chain that comprises the amino acid sequences of L-CDR1, L-CDR2 and L-CDR3, of the MOR-7480.1, MOR-7480 or MOR-7480.2 antibody, or, as part of the description, an antibody selected from the group consisting of MOR- 6032, MOR-7361, MOR-7483, MOR-7483.1 or MOR-7483.2, or sequences having changes of said CDR sequences selected from the group consisting of conservative changes, in which conservative changes are selected from the group consisting of replacement of non-polar waste with other non-polar waste, replacement of polar charged waste with other polar non-charged waste, replacement of polar charged waste with other polar charged waste, and the replacement of structurally similar waste; non-conservative substitutions, in which the non-conservative substitutions are selected from the group consisting of the substitution of the polar charged residue with the uncharged polar residues and the substitution of the non-polar residues with the polar residues, additions and deletions.

In a further embodiment, the anti-4-1BB antibody contains less than 10, 7, 5 or 3 amino acid changes of the germline sequence in the framework or CDR regions. In another embodiment, the antibody contains less than 5 amino acid changes in the framework regions and less than 10 changes in the CDR regions. In a preferred embodiment, the antibody contains less than 3 amino acid changes in the framework regions and less than 7 changes in the CDR regions. In a preferred embodiment, the changes in the framework regions are conservative and those in the CDR regions are somatic mutations.

In another embodiment, the anti-4-1BB antibody has at least 80%, more preferably, at least 85%, even more preferably, at least 90%, even more preferably, at least 95%, more preferably, in minus 99%, of sequence identity on the CDR-1, CDR-2 and CDR-3 heavy and light chain sequences with the CDR sequences of MOR-7480.1, MOR-7480 or MOR-7480.2, or as part of the description, MOR- 6032, MOR-7361, MOR-7483, MOR-7483.1 or MOR-7483.2. Even more preferably, the antibody shares a 100% sequence identity over the light and heavy chain CDR-1, CDR-2 and CDR-3 sequences with the CDR sequences of MOR-7480.1, MOR-7480 or MOR-7480.2 , or as part of the description M ^or R-6032, MOR-7361, MOR-7483, MOR-7483.1 or MOR-7483.2.

In yet another embodiment, the anti-4-1BB antibody has at least 80%, more preferably, at least 85%, even more preferably, at least 90%, even more preferably, at least 95%, more preferably , at least 99%, of sequence identity on the sequences of the variable region of the heavy and light chain with the sequences of the variable region of MOR-7480.1, MOR-7480, MOR-7480.2, or as part of the disclosure MOR-6032, MOR-7361, MOR-7483, MOR-7483.1, or MOR-7483.2. Even more preferably, the antibody shares a 100% sequence identity over the heavy and light chain variable region sequences with the MOR-7480.1 variable region sequences, or as part of the disclosure MOR-6032, MOR- 7361, MOR-7480, MOR-7480.2, MOR-7483, MOR-7483.1 or MOR-7483.2.

II. ADCC inducing antibodies

The embodiments described herein encompass the use of any antibody that induces ADCC. Preferred antibodies that induce ADCC include, but are not limited to, anti-CD20 antibodies (such as rituximab). However, one skilled in the art would appreciate that a variety of antibodies that induce ADCC could be used in the description of this document. For example, suitable antibodies can be modified to have greater ADCC inducing activity (see, for example, Lazar et al., PNAS 2006; 103; 4005-4010; Bowles et al., Blood 2006; 108: 2648-2654 ; Shields et al., JBC 2002; 277: 26733-26740; Suzuki et al., Clin Cancer Res 2007; 13 (6); and Satoh et al., Expert Opin. Biol. Ther. 2006; 6 (11): 1161-1173). Therefore, for example, anti-CD20 antibodies, such as rituximab, could be modified according to methods known in the art to produce ADCC antibodies suitable for use in the embodiments described herein. Similarly, anti-P-cadherin antibodies, such as g-194-g09 (described herein), could be modified according to methods known in the art to produce ADCC antibodies suitable for use in the disclosure herein. document.

The description herein also refers to antibodies that have the amino acid sequences of the CDRs of the heavy and light chains of rituximab and g-194-g09. In other embodiments, the ADCC-inducing antibody is selected from an antibody that has the full length, variable region, or CDR, amino acid sequences of the heavy and light chains of rituximab.

Although the anti-4-1BB antibodies and ADCC inducing antibodies discussed herein may be preferred, those skilled in the art, based on the description provided herein, would appreciate that the embodiments encompass a wide variety of anti antibodies. -4-1BB and ADCC-inducing antibodies and is not limited to the particular antibodies described herein. More particularly, though typically human antibodies are preferred, embodiments are not limited in any way to human antibodies; rather, the embodiments encompass useful antibodies regardless of the origin of the species and include, among others, humanized and / or primed chimeric antibodies. Embodiments include anti-4-1BB antibodies and ADCC-inducing antibodies produced by any method, including, but not limited to, a method known in the art (eg, screening of phage display libraries, and the like) or that are will develop in the future to produce an anti-4-1BB antibody of the invention.

The present embodiments encompass human antibodies produced using a non-human transgenic mammal, that is, XenoMouseMR (Abgenix, Inc., Fremont, CA) as described in U.S. 6,682,736, from Hanson et al.

Another transgenic mouse system for the production of "human" antibodies is known as "HuMAb-MouseMR" (Medarex, Princeton, NJ), which contains miniloci of the human immunoglobulin gene encoding non-rearranged heavy chain human immunoglobulin sequences (mu and gamma) and kappa light chain, together with targeted mutations that inactivate the endogenous mu and kappa chain loci (Lonberg et al. Nature 368: 856-859 (1994), and U.S. Patent No. 5,770,429) .

However, the embodiments encompass antibodies produced using any transgenic mammal such as, but not limited to, the Kirin TC MouseMR (Kirin Beer Kabushiki Kaisha, Tokyo, Japan) as described in, for example, Tomizuka et al., Proc Natl. Acad Sci USA 97: 722 (2000); Kuroiwa et al., Nature Biotechnol 18: 1086 (2000); Publication of United States Patent Application No. 2004/0120948, by Mikayama et al. ; and e1HuMAb-MouseMR (Medarex, Princeton, NJ) and XenoMouseMR (Abgenix, Inc., Fremont, CA), cited above. Therefore, the invention encompasses the use of an antibody produced using any transgenic animal or other non-human animal.

In another embodiment, the antibodies employed in the procedures described herein are not completely human, but "humanized." In particular, murine antibodies or antibodies of other species can be "humanized" or "primatized" using techniques well known in the art. See, for example, Winter and Harris Immunol. Today 14: 43-46 (1993), Wright et al. Crit. Reviews in Immunol. 12: 125-168 (1992), and U.S. Patent No. 4,816,567, to Cabilly et al., And Mage and Lamoyi in Monoclonal Antibody Production Techniques and Applications pages 79-97, Marcel Dekker, Inc., New York, NY (1987).

As will be appreciated based on the description provided herein, the antibodies for use herein can be obtained from a non-human transgenic mammal, and from hybridomas derived therefrom, but can also be expressed on cell lines other than hybridomas. .

Mammalian cell lines available as hosts for expression are well known in the art and include many immortalized cell lines available through the American Type Culture Collection (ATCC), which include, among others, Chinese hamster ovary (CHO) cells ), NSO (also called NS0), HeLa cells, baby hamster kidney cells (BHK), monkey kidney cells (COS) and human hepatocellular carcinoma cells (for example, Hep G2). Prokaryotic and non-mammalian eukaryotic cells, including bacterial, yeast, insect and plant cells, can also be used.

Various expression systems well known in the art can be used, such as, but not limited to, those described in, for example, Sambrook and Russell, Molecular Cloning, A Laboratory Approach, Cold Spring Harbor Press, Cold Spring Harbor, NY ( 2001), and Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, NY (2002). These expression systems include systems based on dihydrofolate reductase (DHFr), among many others. The glutamine synthetase expression system is fully or partially discussed in connection with European patents numbers EP 216846, EP 256055 and EP 323997 and European patent application 89303964. In one embodiment, the antibody used is manufactured in cells. NS0 using a glutamine synthetase system (GS-NS0). In another embodiment, the antibody is manufactured in CHO cells using a DHFR system. Both systems are well known in the art and are described, among others, in Barnes et al. Biotech & Bioengineering 73: 261-270 (2001), and references cited therein.

Site-directed mutagenesis of the CH2 domain of the antibody to eliminate glycosylation may be preferred to prevent changes in immunogenicity, pharmacokinetics and / or effector functions resulting from non-human glycosylation. In addition, the antibody can be deglycosylated by enzymatic procedures (see, for example, Thotakura et al. Meth. Enzymol. 138: 350 (1987)) and / or chemical (see, for example, Hakimuddin et al., Arch. Biochem. Biophys .259: 52 (1987)).

In addition, the embodiments encompass the use of an antibody comprising an altered glycosylation pattern. The person skilled in the art would appreciate, based on the description provided herein, that an antibody can be modified to comprise additional, lesser, or different glycosylation sites compared to the naturally occurring antibody. Such modifications are described, for example, in the publications of United States patent applications Nos. 2003/0207336 and 2003/0157108, and in international patent publications numbers WO 01/81405 and 00/24893.

In addition, some embodiments comprise the use of one or more antibodies independently of the glycoform, if present, present in the antibody. In addition, the methods for extensively remodeling the glycoform present in a glycoprotein are well known in the art and include, for example, those described in international patent publications numbers WO 03/031464, WO 98/58964 and W ^or 99/22764, and publications of United States patent applications Nos. 2004/0063911, 2004/0132640, 2004/0142856, 2004/0072290, and United States Patent No. 6,602,684 to Umaña et al.

In addition, the embodiments encompass the use of an antibody with any covalent and non-covalent modification known in the art, which includes, but is not limited to, binding the polypeptide to one of a variety of non-protein polymers, for example, polyethylene glycol, polypropylene glycol. or polyoxyalkylenes in the manner set forth in, for example, the patent application publications of e E. Uu. Nos. 2003/0207346 and 2004/0132640, and United States Patents Numbers 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; 4,179,337.

Additionally, the embodiments encompass the use of an antibody, or an antigen binding portion thereof, a chimeric protein comprising, for example, a human serum albumin polypeptide, or a fragment thereof. If the chimeric protein is produced using recombinant methods, for example, by cloning a chimeric nucleic acid encoding the chimeric protein, or by chemical bonding of the two peptide portions, the person skilled in the art will understand once armed with the teachings provided herein that such chimeric proteins are well known in the art and may confer desirable biological properties such as, but not limited to, greater stability and half-life in serum for the antibody of the invention and, therefore, such molecules. They are included in this document.

Antibodies that are generated for use in the invention need not initially possess a particular desired isotype. Rather, the antibody that is generated can possess any isotype and can be subsequently changed using conventional techniques. These include direct recombination techniques (see, for example, U.S. Patent No. 4,816,397) and cell-cell fusion techniques (see, for example, U.S. Patent No. 5,916,771).

The effector function of the antibodies of the invention can be changed by changing the isotype to an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE or IgM for various therapeutic uses. In addition, the dependence of the complement for cell destruction can be avoided by the use of bispecifics, immunotoxins or radiomarkers, for example.

Preferably, the anti-4-1BB antibody is MOR-7480.1 and the ADCC inducing antibody is rituximab.

III. Combination therapy with anti-4-1BB antibodies and antibodies capable of inducing ADCC

The embodiments described herein refer to a combination therapy comprising the co-administration of an anti-4-1BB antibody (for example, MOR-7480.1, MOR-7480 or Mo R-7480.2, or as part of the disclosure MOR-6032, MOR-7361, MOR-7483, m Or-7483.1, or MOR-7483.2) and at least one ADCC-inducing antibody (for example, an anti-CD20 such as rituximab).

In one embodiment, a combination of an anti-4-1BB antibody (for example, MOR-7480.1) and an ADCC inducing antibody is co-administered to a patient to treat cancer. The combination may be useful for the treatment of, among other things, abnormal cell growth, for example, hematological malignancies, B-cell malignant neoplasms, non-Hodgkin lymphoma, non-Hodgkin lymphoma positive for CD20, mesothelioma, hepatobiliary (hepatic duct and biliary), a primary or secondary CNS tumor, a primary or secondary brain tumor, lung cancer (NSCLC and SCLC), bone cancer, pancreas cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, ovarian cancer, colon cancer, rectal cancer, anal region cancer, stomach cancer, gastrointestinal (gastric, colorectal and duodenal), breast cancer (e.g., triple negative breast cancer), uterine cancer, carcinoma of fallopian tubes, endometrial carcinoma, cervical carcinoma, carcinoma of the vagina, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small bowel cancer, canker er of the endocrine system, thyroid gland cancer, parathyroid gland cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, testicular cancer, chronic or acute leukemia, leukemia chronic myeloid, lymphocytic lymphomas, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvis carcinoma, central nervous system (CNS) neoplasms, CNS primary lymphoma, non-Hodgkin lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocortical cancer, gallbladder cancer, multiple myeloma, cholangiocarcinoma, fibrosarcoma, neuroblastoma, retinoblastoma or a combination of one or more of the previous cancers.

In some embodiments, a combination of an anti-4-1BB antibody (eg, MOR-7480.1) and rituximab is co-administered to a patient to treat hematologic malignancies, B-cell malignant tumors, non-Hodgkin lymphoma or non-Hodgkin positive lymphoma. for CD20.

In some description, a combination of an anti-4-1BB antibody (for example, MOR-7480.1) and an anti-P-cadherin antibody is co-administered to a patient to treat breast cancer (eg, triple breast cancer negative).

In addition, the invention encompasses the use of an anti-4-1BB antibody in combination with an anti-CD20 antibody such as rituximab as a neoadjuvant, adjuvant, first line treatment, second line therapy and / or third line for cancer. (for example, adjuvant therapy for breast cancer, first-line therapy for metastatic lung cancer, third-line therapy for germ cell tumors and the like).

In one embodiment, the combination of the invention is administered in additional combination with a standard care therapy for one of the cancers described above. In another embodiment, the combination is administered to a patient who has failed standard care therapy.

The person skilled in the art would appreciate, once provided the teachings described herein, that the procedures described herein can be used with, or sequentially (before or after) with surgery, radiotherapy, or both, to treat cancer. . That is, several treatments can be combined with antibody combination therapy, as one skilled in the art would understand once armed with the teachings provided herein.

In another embodiment, an anti-4-1BB antibody in combination with at least one ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab) is co-administered to enhance, prolong, or both, an immune response to a tumor. This is because there may be an interaction between the anti-tumor effect of the anti-4-1BB antibody and the ADCC inducing antibody that leads to a more effective anti-tumor effect than any of the antibodies alone. Therefore, without wishing to be bound by any particular theory, the combination of the anti-4-1BB antibody and the ADCC-inducing antibody can induce a more robust immune response within the tumor than expected. Therefore, the combination of the anti-4-1BB antibody and the ADCC inducing antibody can provide a possible additive or synergistic effect, thus providing an important novel therapeutic treatment for cancer.

In certain embodiments, the ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab) can enhance the effects of the anti-4-1BB antibody in an additive manner. In a preferred embodiment, the ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab) potentiates the effects of the anti-4-1BB antibody in a synergistic manner. In another embodiment, the anti-4-1BB antibody enhances the effect of an ADCC inducing antibody in an additive manner. Preferably, the effects are synergistically enhanced. Therefore, in certain embodiments, the invention encompasses disease treatment or prevention methods that provide better therapeutic profiles than the administration of the ADCC-inducing antibody (eg, an anti-CD20 antibody such as rituximab) alone and / or anti-antibody. 4-1BB alone.

The present embodiments encompass combination therapies that have additive potency or an additive therapeutic effect, while reducing or avoiding adverse or unwanted effects. The invention also encompasses synergistic combinations in which the therapeutic efficacy is greater than the additive, while adverse or undesirable effects are reduced or avoided. In certain embodiments, the methods of the invention allow the treatment or prevention of diseases and disorders in which the treatment is improved by an improved antitumor response using lower and / or less frequent doses of anti-4-1BB antibody and / or ADCC inducing antibody to reduce the incidence of unwanted or adverse effects caused by the administration of anti-4-1BB antibody and / or ADCC inducing antibody alone, while maintaining or improving the efficacy of the treatment, preferably increases patient compliance , improves therapy and / or reduces unwanted or adverse effects.

The methods and compositions of the invention are useful not only in untreated patients but also are useful in the treatment of patients who do not partially or completely respond to the ADCC-inducing antibody administered alone or to the anti-4-1BB antibody administered alone. Various embodiments provide methods and compositions useful for the treatment of diseases or disorders in patients that have proven to be or may be refractory or that do not respond to therapies that comprise the administration of one or both anti-4-1BB antibodies and / or that induce ADCC , and in which the treatment is improved by an improved immune response. In one embodiment, the method comprises combining an anti-CD20 antibody (preferably, rituximab) and an anti-4-1BB antibody (preferably, MOR-7480.1 antibody).

The antibodies can be administered in doses, compositions, by dosing regimens and by the routes of administration described herein. The person skilled in the art would appreciate, based on the description provided herein, that the dose and dosage regimen are adjusted according to procedures well known in the therapeutic techniques. That is, the maximum tolerable dose can be easily established, and the effective amount that provides a detectable therapeutic benefit to a patient can also be determined, as well as the temporary requirements to administer each agent to provide a detectable therapeutic benefit to the patient. Accordingly, although certain dosage and administration regimens are exemplified herein, these examples do not limit the dose and dosage in any way. administration regimen that can be provided to a patient when practicing the present invention. In addition, one skilled in the art would understand, once armed with the teachings provided herein, that a therapeutic benefit, such as, among others, a detectable decrease in tumor size and / or metastasis, a reduced level of PSA In prostate cancer, and the recurrence time, among many other parameters, can be assessed by a wide variety of procedures known in the art to evaluate the efficacy of cancer treatment, and these procedures are included herein, as well as the procedures that will be developed in the future.

Some embodiments described herein refer to neoadjuvant, adjuvant, first-line and / or second-line therapy comprising administering a combination of antibodies. Other embodiments include the use of the combination throughout the disease and continuous treatment. More specifically, the new procedures described herein may provide a therapeutic benefit before and after metastasis, as well as to patients who have become refractory to a chemotherapeutic agent, since the combination of antibodies can improve an immune response, including any response mediated by therapy. The data disclosed herein suggests that immunotherapy comprising an anti-4-1BB antibody in combination with at least one ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab or an anti-P-cadherin antibody) It can provide a therapeutic benefit alone or in combination with at least one additional agent, at any time during treatment. In fact, the data disclosed herein further suggest that a synergistic effect is mediated by the combined administration of an anti-4-1BB antibody and at least one ADCC inducing antibody (eg, an anti-CD20 antibody such as the rituximab or an anti-P-cadherin antibody) for the treatment of cancer. Therefore, the present embodiments provide important novel therapies for the treatment of cancer, whereby the patient's immune system is improved to provide an antitumor effect.

IV. Additional combination therapy

Based on the description provided herein, which includes the combined additive or synergistic effect of the co-administration of an anti-4-1BB antibody in combination with an ADCC-inducing antibody (eg, an anti-CD20 antibody such as rituximab), the person skilled in the art would appreciate that the invention encompasses numerous combination therapies in which the antibodies are administered to the patient in combination with at least one other therapeutic agent that provides a therapeutic benefit. Although many of these combinations will be readily apparent to one skilled in the art once armed with the teachings provided in this document, several combinations are discussed in this document. However, the embodiments described herein are not limited in any way to these combinations, which are set forth herein for illustrative purposes only.

The co-administration of the antibodies with an additional therapeutic agent encompasses the co-administration of the anti-4-1BB antibody, an ADCC-inducing antibody (eg, an anti-CD20 antibody such as rituximab) and one or more additional therapeutic treatments, and also it encompasses the co-administration of two or more separate pharmaceutical compositions, one comprising the anti-4-1BB antibody and the other or others comprising the ADCC inducing antibody, and another or others comprising at least one additional therapeutic agent. In addition, although joint or combined (joint) administration therapy generally means that antibodies and additional therapeutic agents are administered at the same time, it also encompasses simultaneous, sequential or separate dosing of the individual components of the treatment. In addition, when an antibody is administered intravenously and the additional therapeutic agent (s) are administered orally, or by subcutaneous or intramuscular injection, it is understood that the combination is preferably administered as two, three or more separate pharmaceutical compositions.

When a mammal undergoes additional chemotherapy, chemotherapeutic agents well known in the art can be used in combination with the procedures described herein. In addition, growth factor inhibitors, biological response modifiers, alkylating agents, intercalating antibiotics, vinca alkaloids, immunomodulators, taxanes, selective estrogen receptor modulators (SERMs), such as , angiogenesis inhibitors, lasofoxifene, among many therapeutic agents, some of which are described below, can be used.

Angiogenesis inhibitors

An angiogenesis inhibitor can be used in the procedures described herein. An angiogenesis inhibitor includes, but is not limited to, bevacizumab (AVASTIN; Genentech), a humanized antibody against VEGF. It can be used in combination with 5FU and is indicated as a first-line treatment for patients with metastatic carcinoma of the colon or rectum. Agents that directly attack the angiogenic factors or their receptors offer the possibility of increased activity in the competent hematologic neoplasms of the recipient by interrupting the signaling of the autocrine receptor. Bevacizumab produces a sustained neutralization of circulating VEGF and may be useful for the treatment of myelodysplastic syndrome (MDS), lymphoma, acute myeloid leukemia ( ^a M ^l ) and solid tumors. Small molecule RTKI inhibitors of angiogenic receptor signaling (eg, indolinone) are included in the embodiments. described in this document. The first receptor antagonist that enters clinical trials in hematologic malignancies is SU5416 (Sugen), which affects ligand-induced autophosphorylation of VEGFR-1 and VEGFR-2 and c-Kit receptors. SU5416 inhibits the VEGF-induced clonogenic response in leukemia cell lines and promotes apoptosis in myeloblasts of patients with AML. Other RTKIs are being evaluated, including PTK787 / ZK222584 (Novartis) and AG-13736 (Agouron / Pfizer) to treat AML and other hematologic malignancies competent for the recipient. The embodiments described herein also include the treatment of cancer, for example, lymphomas, colon carcinomas, renal carcinoma, gastrointestinal stromal tumors and the like, using a combination of an anti-4-1BB antibody, at least one inducing antibody of ADCC (for example, an anti-CD20 antibody such as rituximab), and at least one additional angiogenesis inhibitor, for example, AG-13736, ^to G-26,798, and the like, as well as other angiogenesis inhibitors that are well known in the art or developed in the future.

Thus, antiangiogenic agents, such as MMP-2 inhibitors (matrix metalloproteinase 2), MMP-9 inhibitors (matrix metalloproteinase 9) and COX-II inhibitors (cyclooxygenase II), can be used in the procedures described in this document. Examples of useful COX-II inhibitors include CELEBREXMR (celecoxib), valdecoxib, rofecoxib, parecoxib, deracoxib, SD-8381, ABT-963, etoricoxib, lumiracoxib, BMS-347070, NS-398, RS 57067, meloxicam. Examples of useful matrix metalloproteinase inhibitors are described in international patent publications numbers WO 96/33172; ^w O 96/27583; WO 98/07697, WO 98/03516, WO 98/34918, WO 98/34915, WO 98/33768, WO 98/30566, WO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667, European Patent Applications Nos. 780386 (published June 25, 1997), 97304971.1 (filed July 8, 1997), 99308617.2 (filed October 29, 1999), 606046 (published July 13, 1994) , 931788 (published July 28, 1999), 99302232.1 (filed March 25, 1999), international application PCT / IB98 / 01113 (filed July 21, 1998), Britain's patent application No.

9912961.1 (filed June 3, 1999), provisional U.S. patent application No. 60 / 148,464 (filed August 12, 1999), and U.S. patents Nos. 5,863,949 and 5,861,510.

Preferred MMP-2 and MMP-9 inhibitors are those that have little or no MMP-I inhibitory activity. More preferred are those that selectively inhibit MMP-2 and / or MMP-9 in relation to the other matrix metalloproteinases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, M ^m P-6, MMP-7, MMP-8, MMP-10, MMP-I I, MMP-12 and MMP-13).

Inhibitors of signal transduction.

The treatments described herein can also be used with other signal transduction inhibitors, such as agents that can inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, antibodies. EGF and molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors, such as VEGF receptors and molecules that can inhibit VEGF; and erbB2 receptor inhibitors, such as organic molecules or antibodies that bind to the erbB2 receptor, for example, HERCEPTIN (Genentech, Inc., San Francisco, CA).

EGFR inhibitors are described, for example, in international patent publications Nos WO 95/19970, WO 98/14451, WO 98/02434 and in U.S. Patent No. 5,747,498, and such substances can be used in the present invention as described herein. EGFR inhibitors include, but are not limited to, monoclonal antibodies C225 (ERBITUX), 22Mab anti-EGFR (ImClone Systems Inc., New York, NY) and ABX-EGF (panitumumab, Abgenix Inc., Fremont, CA ), compounds ^z D-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX-447 (Medarex, lnc., Annandale, NJ) and OlX-103 (Merck & Co., Whitehouse Station, NJ), VRCTC -310 (Ventech Research) and EGF fusion toxin (Seragen Inc., Hopkinton, MA). These and other EGFR inhibitors can be used in the present embodiments.

Compounds directed to the inhibition of tyrosine kinase (TK) of the epidermal growth factor receptor (EGFR) represent a relatively new class of antineoplastic drugs that are useful in the process of the present invention. Many human cancers express members of the EGFR family on the cell surface. When a ligand binds to EGFR, it triggers a cascade of cellular reactions that result in an increase in cell division and influence other aspects of cancer development and progression, including angiogenesis, metastatic spread and inhibition of apoptosis. EGFR-TK inhibitors can selectively target one of the members of the EGFR family (EGFR (also known as HER1 or ErbB-1), HER2 / neu (also known as ErbB-2), HER3 (also known as ErbB- 3), or HER4 (also known as ErbB-4)), or you can point to two or more of them. Suitable EGFR-TK inhibitors for use in the present invention include gefitinib (IRESSA), erlotinib (TARCEVA), CI-1033 (Pfizer), GW2016 (GlaxoSmithKline), EKB-569 (Wyeth), PKI-166 (Novartis), CP-724,714 (Pfizer), and BIBX-1382 (Boehringer-Ingelheim). Additional EGFR-TK inhibitors are described in US patent application number 09 / 883,752, filed June 18, 2001. VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc., San Francisco , CA), can also be used in combination with the procedures described herein. VEGF inhibitors are described, for example, in International Patent Application No. PCT / IB99 / 00797 (filed May 3, 1999), International Patent Publications Nos. Wo 99/24440; WO 95/21613; WO 99/61422; WO 98/50356; WO 99/10349; WO 97/32856; WO 97/22596; WO 98/54093; WO 98/02438; WO 99/16755; WO 98/02437; United States patents Nos. 5,834,504; 5,883,113; 5,886,020; and 5,792,783. Other examples of some VEGF specific inhibitors useful in the present invention are IM862 (Cytran Inc., Kirkland, WA); Imclone IMC-1C11 antibody, anti-VEGF monoclonal antibody from Genentech, Inc., San Francisco, CA; and angiozyme, a synthetic ribozyme from Ribozyme (Boulder, CO) and Chiron (Emeryville, CA).

ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc) and monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc., Woodlands, TX) and 2B-1 (Chiron), can also be combined with the procedures described herein, by example, those indicated in international patent publications Nos. WO 98/02434; WO 99/35146; WO 99/35132; WO 98/02437; WO 97/13760; WO 95/19970; United States patents Nos.

5,587,458 and 5,877,305. ErbB2 receptor inhibitors useful in the present embodiments are also described in EP1029853 (published August 23, 2000) and in International Patent Publication No. WO 00/44728 (published August 3, 2000). The compounds and substance that inhibit the erbB2 receptor described in the PCT applications mentioned above, United States patents, and provisional applications of the United States, as well as other compounds and substances that inhibit the erbB2 receptor, can be used with antibodies in accordance with the present embodiments.

The treatments described herein can also be used with other agents useful for treating abnormal cell growth or cancer, which include, among others, other agents capable of improving antitumor immune responses; and antiproliferative agents such as farnesyl transferase protein inhibitors (eg, BMS 214662), and avp3 inhibitors, such as the VITAXlN antibody of avp3, avp5 inhibitors, p53 inhibitors and the like.

When the antibodies of the methods described herein are administered in combination with another immunomodulatory agent, the immunomodulatory agent may be selected, for example, from the group consisting of a dendritic cell activator such as the CD40 ligand and the anti-CD40 agonist antibodies. as well as antigen presentation enhancers, T cell tropism enhancers, tumor-related immunosuppressive factor inhibitors, such as TGF-p (transforming growth beta factor) and IL-10. Preferred anti-CD40 agonist antibodies encompass the antibodies described in International Patent Application No. PCT / US02 / 36107, filed on November 8, 2002 (published as WO 03/040170 on May 15, 2003), and the application U.S. Patent No. 10 / 292,088, filed on November 8, 2002 (published as U.S. Patent Publication No. US2003 / 0211100 on November 13, 2003), which includes, among others, an antibody that It has the amino acid sequence of the heavy and light chains of antibody 3.1.1, 3.1.1.H-A78T, 3.1.1H-A78T-V88A-V97A, 3.1.1L-L4M-L83V, 3.1.1H-A78T-V88A -V97A / 3.1.1L-L4M-L83V, 7.1.2, 10.8.3, 15.1.1, 21.2.1, 21.4.1, 22.1.1, 22.1.1H-C109A, 23.5.1, 23.25.1, 23.28 .1, 23.28.1H-D16E, 23.29.1 and 24.2.1.

IGF-1R inhibitors

The treatment regimens present can also be combined with antibodies or other ligands that inhibit tumor growth by binding to IGF-1R (insulin-like growth factor 1 receptor). Examples of anti-IGF-1R antibodies that can be used include those described in International Patent Application No. PCT / US01 / 51113, filed on 12/20/01 (published as WO 02/053596 on July 11, 2002 ), and international patent application No. PCT / IB2004 / 002555, filed on August 3, 2004 (published as WO 2005/016967 on February 24, 2005). Preferred anti-IGFR-1R antibodies encompass an antibody having the amino acid sequence of the heavy and light chains, for example, of the antibody 2.12.1, 2.13.2, 2.14.3, 3.1.1, 4.9.2 and 4.17 .3.

Ligands that inhibit signaling through IGF-1R also encompass small molecules and other ligands that include, among others, somavert (PEGVISOMANT), which is a growth hormone analog that inhibits IGF-1 signaling. PEGVISOMANT is conjugated with polyethylene glycol and can be used, among other things, to treat acromegaly. PEGVISOMANT can be co-administered with antibodies from the procedures described herein to treat cancer, since the combination can inhibit tumor growth. Therefore, PEGVISOMANT, similarly with anti-IGF-1R antibodies, can be used to treat cancer as described herein.

The present embodiments encompass therapies that can also be combined with a large number of therapeutic, surgical, radiation and other therapeutic therapies to treat a patient. The therapeutic agents are numerous and have been described, for example, in US Patent Application Publications Nos.

2004/0005318, 2003/0086930, 2002/0086014, and the international publication No. WO 03/086459, among many others. Such therapeutic agents include, but are not limited to, topoisomerase I inhibitors; other antibodies (trastuzumab, anti-IGF-1R and the like); chemotherapeutic agents such as imatinib (GLEEVEC, GLIVEC or STI571; Novartis), sorafenib (BAY 43-9006; Bayer Pharmaceuticals Corp./Onyx Pharmaceuticals), selective estrogen receptor modulators (SERM), taxanes, vinca alkaloids , temozolomide, angiogenesis inhibitors, EGFR inhibitors, VEGF inhibitors, ErbB2 receptor inhibitors, antiproliferative agents (eg, farnesyl transferase protein inhibitors and avp3 inhibitors, avp5 inhibitors, p53 inhibitors and the like), immunomodulators , cytokines, tumor vaccines; tumor specific antigens; stem and dendritic cell therapies; alkylating agents, folate antagonists; pyrimidine antagonists; antibiotics anthracycline; platinum compounds; costimulatory molecules (for example, CD4, CD25, PD-1, B7-H3, 4-1BB, OX40, ICOS, CD30, HLA-DR, MHCII and LFA).

Radiotherapy

Radiation therapy can be co-administered with the antibody combination therapies described herein. Radiation therapy is given according to well-known radiation therapy procedures for the treatment of cancer, such as breast cancer. The dose and regimen for radiotherapy can easily be determined by one skilled in the art and based on the stage of the disease and other factors well known in the art.

Palliative Agents

The present embodiments also encompass the administration of other therapeutic agents. Such therapeutic agents include analgesics, cancer vaccines, antivascular agents, antiproliferative agents, antiemetic agents and antidiarrheal agents. Preferred antiemetic agents include ondansetron hydrochloride, granisetron hydrochloride and metoclopramide. Preferred antidiarrheal agents include diphenoxylate and atropine (LOMOTIL), loperamide (IMMODIUM) and octreotide (SANDOSTATINE).

In another embodiment, the methods described herein include administering an agent with antidiarrheal effect in which the agent is indicated in the treatment of chronic inflammatory conditions of the gastrointestinal tract. Such agents include, but are not limited to, steroids with topical activity (for example, budesonide [ENTOCORT]), and drugs against tumor necrosis factor (TNF) (for example, infliximab [REMICADE], etanercept [ ^e N ^b REL] and adalimumab [SHOULD]).

Stem cell-based therapy

The antibody combination therapy described herein can be combined with stem cell transplantation to provide a therapeutic benefit to a cancer patient. Stem cell transplantation can be performed according to procedures known in the art. Some of these procedures are described in Appelbaum in the Harrison's Principles of Internal Medicine, Chapter 14, Braunwald et al., Eds., 15th ed., McGraw-Hill Professional (2001). Therefore, the methods of the present disclosure relate to the treatment of cancer in a mammal that has undergone a stem cell transplant, procedures comprising administering to the mammal an amount of an anti-4-1BB antibody in combination with the less an ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab or an anti-P-cadherin antibody), which is effective in treating cancer in additional combination with stem cell transplantation.

When the procedure comprises stem cell transplantation, the first dose of the antibody combination therapy can be administered after the mammalian immune system has recovered from the transplant, for example, in the period of one to 12 months after the transplant. In certain embodiments, the first dose is administered in the period of one to three, or one to four months after the transplant. The patient can undergo stem cell transplantation and preparatory treatment or treatments.

The procedures described herein also refer to procedures for the treatment of cancer in a mammal comprising the steps of (i) performing a stem cell transplant in the mammal and (ii) administering an effective amount of an anti-antibody. 4-1BB in combination with an effective amount of at least one ADCC inducing antibody. Preferably, the mammal is a human. The stem cell transplant can be allogeneic or autologous stem cell. In addition, cell transplantation encompasses the adoptive transfer of lymphocytes, either from the same patient and / or from an HLA-compatible donor.

In addition, the methods of the disclosure can be combined with radiotherapy and stem cell transplantation, and any combination of any of the treatments described herein, known in the art, or to be developed in the future.

When the combination antibody treatment is combined with a standard cancer treatment, such as, among others, chemotherapeutic regimens, it may be possible to reduce the dose of chemotherapeutic reagent administered (Mokyr, M. et al. Cancer Research 58: 5301 - 5304 (1998)). The combination therapies described herein can provide an increased source of tumor-specific antigens, thus providing an increased immune response to the tumor, which, in turn, provides a therapeutic benefit to the patient.

V. Dosing regimens

For administration of one or more antibodies described herein, the dosage regimens may be adjusted to provide the desired optimal response. For example, a single bolus can be administered, Several divided doses may be administered over time or the dose may be reduced or increased proportionally as indicated by the requirements of the therapeutic situation. It may be especially advantageous to formulate parenteral compositions in the form of a dosage unit to facilitate administration and dosage uniformity. The dosage unit form as used herein refers to physically discrete units suitable as unit dosages for the mammalian subjects to be treated; Each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The specification for unit dosage forms of the invention is typically dictated and depends directly on (a) the unique characteristics of the antibody and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the composition technique such as an active compound for the treatment of sensitivity in individuals.

For administration of antibodies or combinations of antibodies, the dose may range between about 0.0001 and 100 mg / kg, and more generally between 0.01 and 20 mg / kg, of the host's body weight. A non-limiting range, by way of example, for a therapeutically effective amount of an antibody or combination of antibodies administered according to the procedures described herein is at least about 0.01 mg / kg, at least about 0.1 mg / kg, at least about 0.3 mg / kg, at least about 1 mg / kg, at least about 5 mg / kg, at least about 6 mg / kg, at least about 10 mg / kg, at least about 15 mg / kg, or at least about 20 mg / kg. For example, a therapeutically effective amount of antibody or combination of antibodies may vary from about 0.01-15 mg / kg, or for example about 0.03-3 mg / kg, or for example about 0.1-1 mg / kg In addition, a therapeutically effective amount of antibody may range from about 0.1-30 mg / kg, or for example about 0.3-25 mg / kg, or for example about 1 20 mg / kg, or for example about 3- 20 mg / kg, or for example about 5-20 mg / kg, or for example about 10-20 mg / kg, or about 3-15 mg / kg, or about 5-15 mg / kg, or about 10-15 mg / kg

In addition, an example of a dose escalation protocol can be used to determine the maximum tolerated dose (MTD), to evaluate the dose-limiting toxicity (DLT), if any, associated with the administration of the combination therapy described herein. document, and the like, comprises administering increasing doses, such as, but not limited to, approximately 0.01 mg / kg, 0.03 mg / kg, 0.06 mg / kg, 0.1 mg / kg, 0.12 mg / kg, 0.18 mg / kg, 0.24 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 6 mg / kg, 7 mg / kg, 10 mg / kg, 12 mg / kg, 15 mg / kg, or more than 15 mg / kg, or any combination thereof. In some embodiments, successive doses of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 3 mg / kg, 6 mg / kg, 10 mg / kg, 15 mg / kg or 20 mg are administered / kg and the toxicity for the patient is evaluated, if any, and the effectiveness of the treatment, among other parameters. In some embodiments, successive doses of 0.03 mg / kg, 0.06 mg / kg, 0.12 mg / kg, 0.18 mg / kg, 0.24 mg / kg and 0.3 mg / kg are administered and toxicity is evaluated for the patient, if any, as well as for the effectiveness of the treatment, among other parameters. Such studies to determine the toxicity and efficacy of dose regimens are well known in the art.

It should be taken into account that the dosage values may vary with the type and severity of the condition to be relieved, and may include single or multiple doses. It should also be understood that for any particular subject, specific dosage regimens must be adjusted over time according to the individual need and professional judgment of the person administering or supervising the administration of the compositions, and that the intervals of Dosage set forth herein are examples only and are not intended to limit the scope or practice of the claimed composition. The determination of the appropriate doses and regimens for the administration of the antibody are well known in the relevant art and it will be understood that they are understood by the person skilled in the art once the teachings described herein are provided.

In one embodiment, the antibodies are administered in an intravenous formulation as a sterile aqueous solution containing approximately 5 to 20 mg / mL of antibodies, in an appropriate buffer system.

In one embodiment, part of the dose is administered by an intravenous bolus and the rest by infusion of the antibody formulation. For example, an intravenous injection of antibody can be administered as a bolus, and the rest of a predetermined dose of antibody can be administered by intravenous injection. A predetermined dose of the antibodies can be administered, for example, over a period of about an hour and a half to about five hours.

Some embodiments described herein relate to the administration of a combination of an anti-4-1BB antibody and at least one ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab). The person skilled in the art would appreciate that the combination can be administered simultaneously or that the antibodies and various agents can be administered at different times. However, the present embodiments are not limited to these or any particular dosage or administration regimen for administering an anti-4-1BB antibody in combination with at least one ADCC-inducing antibody (eg, an anti-CD20 antibody such as rituximab ). Rather, the optimal dose, route and regimen for antibody administration can be readily determined by a person skilled in the art using well known procedures.

For example, a single dose or multiple doses of each antibody (or combination of antibodies) can be administered. Alternatively, at least one dose, or at least three, six or 12 doses may be administered. Doses can be administered, for example, daily, weekly, every two weeks, monthly, every twenty days, every 25 days, every 28 days, every 30 days, every 40 days, every 50 days, every two months, every 70 days , every 80 days, every three months, every six months or every year, or any other period that provides a therapeutic benefit to the patient as determined by the skilled professional.

In one embodiment, a single bolus injection comprising the anti-4-1BB antibody is administered to a patient intravenously at a dose ranging from about 0.1 mg / kg to 20 mg / kg approximately every twenty-eight days. A dose of an ADCC inducing antibody is administered that first day. In some embodiments, the antibodies are co-administered on the same day as the start of each dose cycle. In other embodiments, the ADCC-inducing antibody is administered at any point during the administration of the anti-4-1BB antibody, or vice versa, and the invention is not limited in any way with respect to the relative administration of the antibodies. Therefore, the ADCC inducing antibody can be administered before, during and / or after administration of the anti-4-1BB antibody.

The combination of antibodies can be administered as a neoadjuvant therapy before surgery, radiotherapy or any other treatment, to sensitize the tumor cells or to confer a therapeutic benefit to the patient. In addition, the combination can be co-administered as neoadjuvant therapy after localized treatment (eg, surgery, radiation, or both).

In addition, the combination can be administered as a second-line therapy, such as, but not limited to, once the first-line therapy has failed. Alternatively, the combination can be administered simultaneously with the first-line therapy or at any time during the first-line therapy, which can be administered after the initial treatment.

The procedures described herein include the administration of a combination of antibodies, with or without additional therapy, which includes, among others, a hormonal agent, radiotherapy and any additional therapeutic agent (chemotherapy, signal inhibition therapy, among others), and the like, as one skilled in the art would appreciate based on the description provided in this document.

The disclosure also refers to an article of manufacture (for example, a dosage form adapted for iv administration) comprising an anti-4-1BB antibody in the amount effective to treat cancer (for example, at least 0.01 mg / kg, at least 0.03 mg / kg, at least 0.06 mg / kg, at least 0.1 mg / kg, at least 0.12 mg / kg, at least 0.18 mg / kg, at least 0.24 mg / kg, at least 0.3 mg / kg, at least 0.5 mg / kg, at least 0.7 mg / kg, at least 1 mg / kg, at least 3 mg / kg, at least 5 mg / kg, at least 10 mg / kg, at least 15 mg / kg or at least 20 mg / kg) and a therapeutically effective amount of at least one ADCC inducing antibody. In certain embodiments, the article of manufacture comprises a container or containers comprising an anti-4-1BB antibody, at least one ADCC inducing antibody (for example, an anti-CD20 antibody such as rituximab), and a label and / or instructions of use to treat cancer.

SAW. Pharmaceutical compositions

The embodiments described herein encompass the preparation and use of pharmaceutical compositions comprising a human anti-4-1BB antibody of the invention as an active ingredient in combination with at least one ADCC-inducing antibody (eg, an anti-antibody CD20 such as rituximab or an anti-P-cadherin antibody). Said pharmaceutical composition may consist of each active ingredient alone, as a combination of at least one active ingredient (for example, an effective dose of an anti-4-1BB antibody, an effective dose of at least one ADCC inducing antibody, such as rituximab ) in a form suitable for administration to a subject, or the pharmaceutical composition may comprise the active ingredient and one or more pharmaceutically acceptable carriers, one or more additional ingredients (active and / or inactive), or some combination thereof.

In some embodiments, one or more antibodies are administered parenterally (for example, intravenously) in an aqueous solution. In other embodiments, one or more antibodies are administered orally in the form of a pill / capsule. However, the person skilled in the art would understand, based on the description provided herein, that the invention is not limited to these, or any other formulations, doses, routes of administration and the like. Rather, the invention encompasses any formulation or method of administering an anti-4-1BB antibody in combination with an ADCC-inducing antibody, which includes, inter alia, administering each agent separately in a different formulation through a route of administration. different (for example, administering an iv antibody, while co-administering the other antibody orally), among many others. Therefore, the following discussion describes several formulations for practicing the procedures described herein that comprise the administration of any anti-4-1BB antibody in combination with an ADCC-inducing antibody, but the present embodiments are not limited to these formulations. , but include any formulation that can be easily determined by an expert in the technique once armed with the teachings provided in this document for use in the procedures described.

The antibodies employed in the embodiments described herein may be incorporated into pharmaceutical compositions suitable for administration to a subject. Typically, the pharmaceutical composition comprises the antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delay agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, trehalose, glycerol, ethanol and the like, as well as combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol or sodium chloride in the composition. Pharmaceutically acceptable substances such as humectants or minor amounts of auxiliary substances such as humectants or emulsifying agents, preservatives or buffers, which improve the shelf life or effectiveness of the antibody or antibody portion.

The antibodies can be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (for example, injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The preferred form depends on the intended mode of administration and the therapeutic application. Typical preferred compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for passive immunization of humans with other antibodies. The preferred mode of administration is parenteral (eg, intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the antibody is administered by infusion or intravenous injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome or other ordered structure suitable for a high concentration of drug. Sterile injectable solutions can be prepared by incorporating the antibody in the required amount in an appropriate solvent with one or a combination of the ingredients listed above, as required, followed by sterilization by filtration. In general, the dispersions are prepared by incorporating the active compound in a sterile vehicle containing a basic dispersion medium and the other required ingredients of those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation procedures are vacuum drying and freeze drying which produces a powder of the active ingredient plus any additional desired ingredient of a previously sterile filtered solution of the same. The adequate fluidity of a solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of injectable compositions can be achieved by including in the composition an agent that delays absorption, for example, monostearate and gelatin salts.

Antibodies can be administered by a variety of procedures known in the art, including, without limitation, oral, parenteral, mucosal, inhalation, topical, buccal, nasal and rectal. For many therapeutic applications, the preferred route / mode of administration is subcutaneous, intramuscular, intravenous or infusion. A needleless injection can be used, if desired. As those skilled in the art will appreciate, the route and / or mode of administration will vary depending on the desired results.

Dosage regimens can be adjusted to provide the desired optimal response. For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be reduced or increased proportionally as indicated by the requirements of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in the form of a dosage unit to facilitate the administration and uniformity of the dosage. The dosage unit form as used herein refers to physically discrete units suitable as unit dosages for the mammalian subjects to be treated; Each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The specification for the unit dosage forms of the invention is dictated by (a) the unique characteristics of the antibody and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the composition technique, such as a active compound for the treatment of sensitivity in individuals.

It should be taken into account that the dosage values may vary with the type and severity of the condition to be relieved, and may include single or multiple doses. It should also be understood that for any particular subject, specific dosage regimens must be adjusted over time according to the individual need and professional judgment of the person administering or supervising the administration of the compositions, and that the intervals of Dosage set forth herein are examples only and are not intended to limit the scope or practice of the claimed composition.

In one embodiment, one or more antibodies are administered in an intravenous formulation as a sterile aqueous solution containing 5 or 10 mg / mL of antibody, with sodium acetate, polysorbate 80 and sodium chloride at a pH ranging from about 5 to 6. Preferably, the intravenous formulation is a sterile aqueous solution containing 5 or 10 mg / mL of antibody, with 20 mM sodium acetate, 0.2 mg / mL polysorbate 80 and 140 mM sodium chloride at pH 5.5 .

In another embodiment of the invention, one or more antibodies are administered in a sterile solution comprising 20 mM histidine buffer, pH 5.5, 84 mg / mL trehalose dihydrate, 0.2 mg / mL polysorbate 80 and 0.1 mg / mL ethylenediaminetetraacetic acid dihydrate. In one aspect, the formulation is packaged in clear glass vials with a rubber stopper and an aluminum seal. In another aspect, the vial contains approximately 20 mg / mL of antibody with a nominal filling of approximately 400 mg per vial.

In one embodiment, part of the dose is administered by an intravenous bolus and the rest by infusion of the antibody formulation. For example, an intravenous injection of 0.01 mg / kg of the bolus-shaped antibody can be administered, and the remainder of a predetermined dose of antibody can be administered by intravenous injection. A predetermined dose of the antibody can be administered, for example, over a period of one and a half hours up to two hours to five hours.

The formulations of the pharmaceutical compositions described herein can be prepared by any method known or developed below in the art of pharmacology. In general, such preparation procedures include the step of associating the active ingredient with a carrier or one or more additional accessory ingredients, and then, if necessary or desirable, forming or packaging the product in a desired single dose or multi-dose unit. .

A pharmaceutical composition of the invention can be prepared, packaged or sold in bulk, as a single unit dose, or as a plurality of individual unit doses. As used herein, a "unit dose" is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient that would be administered to a subject or a convenient fraction of said dose such as, for example, half or a third of said dose.

The relative amounts of the active ingredient, the pharmaceutically acceptable carrier and any additional ingredient in a pharmaceutical composition will vary, depending on the identity, size and condition of the treated subject and, in addition, depending on the route by which the composition is to be managed. By way of example, the composition may comprise between 0.1% and 100% (w / w) of active ingredient.

In addition to the active ingredient, a pharmaceutical composition of the invention may further comprise one or more additional pharmaceutically active agents. Additional agents particularly contemplated include antiemetics, antidiarrheals, chemotherapeutic agents, cytokines and the like.

Controlled or sustained release formulations of a pharmaceutical composition of the invention can be prepared using conventional technology.

As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by the physical rupture of a tissue of a subject and the administration of the pharmaceutical composition through the rupture in the tissue. Parenteral administration thus includes, but is not limited to, the administration of a pharmaceutical composition by injection of the composition, by applying the composition through a surgical incision, by applying the composition through a non-surgical wound. that penetrates the tissue, and the like. In particular, it is contemplated that parenteral administration includes, among others, subcutaneous, intraperitoneal, intramuscular, intrasternal infusion and renal dialytic infusion techniques.

Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations can be prepared, packaged or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations can be prepared, packaged or sold as a unit dose, such as in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes and implantable sustained-release or biodegradable formulations as described below. Such formulations may further comprise one or more additional ingredients that include, but are not limited to, suspending, stabilizing or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry form (i.e., powdered or granulated) for reconstitution with a suitable vehicle (eg, sterile pyrogen-free water) before parenteral administration of the reconstituted composition

A composition of the present invention can be administered by a variety of procedures known in the art. The route and / or the mode of administration vary according to the desired results. The active ingredients can be prepared with vehicles that protect the active ingredient against rapid release, such as a controlled release formulation, which includes implants, transdermal patches and microencapsulated delivery systems. Biodegradable and biocompatible polymers, such as vinyl ethylene acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid, can be used. Many procedures for the preparation of such formulations are described, for example, in Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, (1978). The pharmaceutical compositions are preferably manufactured under GMP conditions.

The pharmaceutical compositions can be prepared, packaged or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated in accordance with the known technique, and may comprise, in addition to the active ingredient, additional ingredients such as dispersing agents, wetting agents or suspending agents described herein. Such sterile injectable formulations may be prepared using a parenterally acceptable non-toxic diluent or solvent, such as water or 1,3-butanediol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution and fixed oils such as mono or synthetic di-glycerides. Other parenterally administrable formulations that are useful include those comprising the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer system. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials, such as an emulsion, an ion exchange resin, a poorly soluble polymer or a poorly soluble salt.

The combination of the active ingredient of the anti-4-1BB antibody / ADCC inducing antibody can be administered to an animal, preferably a human. The precise administered dose of each active ingredient will vary depending on any number of factors, including, but not limited to, the type of animal and the condition of the disease being treated, the age of the animal and the route or routes of administration.

The anti-4-1BB antibody can be administered to an animal as frequently as several times a day, or it can be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. The frequency of the dose will be readily apparent to the person skilled in the art and will depend on any number of factors, such as, among others, the type and severity of the disease being treated, the type and age of the animal, etc.

The ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab) can be administered to an animal as often as several times a day, or it can be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. The dose frequency will be readily apparent to the person skilled in the art and will depend on any number of factors, such as, among others, the ADCC-inducing antibody itself, as well as the type and severity of the disease being treated, the type and age of the animal, etc.

The anti-4-1BB antibody and the ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab) can be co-administered, since they can be administered separately, at different dates or at different times of the day, as well as simultaneously or on the same date. Therefore, joint administration encompasses any temporary combination of antibody administration, so that the administration of the two mediates a therapeutic benefit to the patient that is markedly greater than the administration of any agent in the absence of the other.

A combination of anti-4-1BB antibody and ADCC inducing antibody can be co-administered with numerous other compounds (antihormonal therapy agents, cytokines, chemotherapeutic and / or antiviral drugs, among many others). Alternatively, the compound or compounds may be administered one hour, one day, one week, one month or even more, before the combination of the anti-4-1BB antibody and the ADCC inducing antibody, or any permutation thereof. In addition, the compound or compounds may be administered one hour, one day, one week or even more, after administration of radiation, stem cell transplantation or administration of any therapeutic agent (e.g., cytokines, chemotherapeutic compounds and the like), or any permutation thereof. The frequency and administration regime will be readily apparent to the person skilled in the art and will depend on any number of factors such as, among others, the type and severity of the disease to be treated, the age and health status of the animal, the identity of the compound or compounds that are administered, the route of administration of the various compounds, and the like. Several instructive examples are provided that demonstrate the methods of co-administration of an anti-4-1BB antibody and an ADCC inducing antibody to treat cancer, but the invention is not limited in any way to these examples, which simply serve to illustrate the procedures covered by the invention.

VII. Kits

Additional embodiments described herein include various kits for cancer treatment. The kits typically include a therapeutically effective amount of a human anti-4-1BB antibody of the invention and a therapeutically effective amount of at least one ADCC-inducing antibody (for example, an anti-CD20 antibody such as rituximab or an anti-P antibody -cadherina). The kit may also include an applicator, which includes, among others, a syringe, for the administration of the kit components to a patient. In addition, the kit may include instructional materials that establish information relevant to the use of the kit to treat cancer in the patient. In addition, the kit may include a large number of additional agents for the treatment of cancer. Such agents are set forth herein and include chemotherapeutic compounds, cancer vaccines, signal transduction inhibitors, agents useful for treating abnormal cell growth or cancer, antibodies or other ligands that inhibit tumor growth, a chemotherapeutic agent ( taxane, vinca alkaloid, platinum compound, interspersed with antibiotics, among many others), and cytokines, among many others, as well as palliative agents to treat, for example, any toxicity that arises during treatment, such as, among others , an antidiarrheal, an antiemetic and the like. Although examples of kits are described herein, the content of other useful kits will be apparent to those skilled in the art in light of the present disclosure.

In some embodiments, the kit comprises at least one anti-4-1BB antibody selected from MOR-6032, MOR-7361, MOR-7480, MOR-7480.1, MOR-7480.2, MOR-7483, MOR-7483.1, MOR -7483.2. In some embodiment, the kit comprises MOR-7480.1 and rituximab. In further embodiments, the kit comprises MOR-7480.1 and anti-P-cadherin1. In further embodiments, the kit comprises MOR-7480.1 and anti-P-cadherin2. Although the kits described above are preferred, the invention is not limited to these particular combinations.

The invention is further described in detail by reference to the following experimental examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise specified. Therefore, the invention should not be construed in any way as limited to the following examples, but should be construed to encompass any and all variations that are apparent as a result of the teachings provided herein.

Examples

Example 1: Combinatorial effects of ADCC-inducing mAb (anti-CD20 mAb) with 4-1BB agonist mAb (MAB9371) in a transplantable tumor model in mice

The combinatorial efficacy of 4-1BB and CD20 substitute mAbs in a lymphoma model was tested. Experiments of tumor growth inhibition were performed in the A20 B-cell lymphoma model, which expresses the B-cell CD20 protein, testing the efficacy of mAb (MAB9371 (R&D Systems (Minneapolis MN)) rat anti-mouse agonist 4-1BB in combination with a mouse mouse anti-CD20 antibody that served as a substitute for rituximab.

One million A20 cells were injected subcutaneously in the right flank of Balb / c mice. The tumors were allowed to grow for 7 days and the animals were randomized according to the tumor volume. A single dose of MAB9371, mouse anti-CD20 or MAB9371 in combination with mouse anti-CD20 was administered i.p. on the day of randomization. MAB9371 was administered in an amount less than optimal at a rate of 0.1 mg / kg or in combination with mouse anti-CD20 at a rate of 5 mg / kg diluted in PBS. A better tumor growth inhibition was obtained when both MAB9371 and CD20 were used together. Tumor size ([length x {width x width}] x 0.5 = volume in mm3) was evaluated every 2-3 days using a digital caliper. The mean ± SEM is shown in Figure 1.

Treatment with the combination increased the efficacy of tumor growth inhibition compared to CD20 or anti 4-1BB alone. In addition, the required dose of 4-1BB in the combination group was reduced and achieved a better inhibition of tumor growth than the dose with anti 4-1BB alone (Figure 1).

Reference Example 2: Combinatorial effects of ADCC-inducing mAb (anti-P-cadherin1 mAb (g-194-g09)) with 4-1BB agonist mAb (MAB9371) in a colon carcinoma model in mice

The combinatorial efficacy of mAbs 4-1BB and P-cadherin was tested in a colon carcinoma model. Tumor growth inhibition experiments were performed, testing the efficacy of the rat mouse anti-4-1BB agonist mAb (MAB9371; R&D Systems (Minneapolis MN)) in combination with an anti-P-cadherin antibody (anti-P-cadherin1) or g-194-g09).

One million CT-26 cells were injected subcutaneously in the right flank of Balb / c mice. The tumors were allowed to grow for 7 days and the animals were randomized according to the tumor volume. A single dose of murine anti-4-1BB (MAB9371), and anti P-cadherin1 or MAB9371 in combination with anti-P-cadherin1 MAB9371 was administered less than optimal at the rate of 0.1 mg / kg or in combination with anti-P-cadherin1 at a rate of 10 mg / kg diluted in PBS. The animals were dosed ip on the day of randomization. Inhibition of Tumor growth was improved by approximately 35% when MAB9371 and anti P-cadherinal (g-194-g09) were used together.

The results show that the 4-1BB mAb (MAB9371), when used in combination with the anticadherin mAb (g-194-g09), improved tumor growth inhibition (Figure 2).

Reference Example 3: Combinatorial effects of ADCC-inducing mAb (anti-P-cadherin2 mAb) with 4-1BB agonist mAb (MAB9371) in a colon carcinoma model in mice

The combinatorial efficacy of mAbs 4-1BB and P-cadherin was tested in a colon carcinoma model. Tumor growth inhibition experiments were performed, testing the efficacy of the rat mouse anti-4-1BB agonist mAb (MAB9371; R&D Systems (Minneapolis MN)) in combination with an anti-P-cadherin antibody (anti-P-cadherin2) ). Anti-P-cadherin2 was modified from anti-P-cadherin1 (g-194-g09) to have a greater ADCC inducing activity.

One million CT-26 cells were injected subcutaneously in the right flank of Balb / c mice. The tumors were allowed to grow for 7 days and the animals were randomized according to the tumor volume. A single dose of murine anti-4-1BB (MAB9371) and enhanced ADCC anti-P-cadherin (anti-P-cadherin2) or MAB9371 in combination with enhanced ADCC anti-P-cadherin was dosed less than optimally at 0.1 mg / kg ratio or in combination with anti-P-cadherin2 at a rate of 10 mg / kg diluted in PBS. The animals were dosed i.p. on the day of randomization. Tumor growth inhibition improved by approximately 35% when both MAB9371 and anti P-cadherin2 were used together.

The results show that mAb 4-1BB (MAB9371) when used in combination with anti-P-cadherin2 mAb, significantly improved tumor growth inhibition (Figure 3).

Example 4: Treatment of animals positive for primary lymphoma with agonist antibody 4-1BB and / or ADCC inducing antibody CD20

The combinatorial efficacy of mAbs 4-1BB and CD20 was tested in Ep-myc mice.

Ep-myc mice (C57BL / 6J-Tg (IghMyc) 22Bri / J) (Jackson Laboratories) were obtained and blood samples were monitored weekly by FACS. Using the established enrollment criteria, the animals were randomly assigned to 6 treatment groups; 1 mg / kg rat IgG2a control qwx2 (Group 1), 1 mg / kg MAB9371 qwx2 (Group 2), 10 mg / kg anti-CD20 of mouse qwx4 (Group 3), 1 mg / kg MAB9371 qwx2 10 mg / kg anti -CD20 mouse qwx4 (group 4), 0.1 mg / kg MAB9371 qwx2 10 mg / kg anti-CD20 mouse qwx4 (group 5), and 1 mg / kg MAB9371 qwx1 10 mg / kg anti-CD20 mouse qwx4 (group 6). Animals continued to be monitored through weekly saphenous vein blood samples and subsequent FACS analysis along with general observation and body weight control. Animals showing obvious signs of disease were sacrificed according to human practices, tissues from groups 1-4 were collected for histopathology and survival was recorded after dosing.

A summary of the groups is shown in Table 3.

-

(continuation)

The mean age at the time of enrollment was 13.6 ± 6.7 weeks for the entire study, without significant differences in the ages between the 6 groups. A significant survival benefit was observed for Group 4 (1 mg / kg MAB9371 qwx2 10 mg / kg anti-CD20 mouse qwx4) compared to Group 1 (1 mg / kg rat IgG2a control qwx2, Table 1) and Group 3 (10 mg / kg anti-CD20 mouse qwx4, (Figure 4, Table 1).

Peripheral blood samples were stained with Fluorochrome-labeled Ab for B220, CD20, CD3, IgM and IgD. The samples were analyzed by flow cytometry using a FACSCanto and FACSDiva software followed by a data analysis using the FlowJo software. Animals that demonstrated an increase in the population of B220lo + above 25% of total lymphocytes combined with an increase in forward dispersion and a week-to-week variation in B220lo of> 5% were randomized to groups of treatment. The day animals were assigned to the group is considered day 0 of the study (n = 10 animals / group). The dosage was based on an average body weight of 22.5 grams / animal. The animals continued to be monitored by flow cytometry, body weight and general observation. Animals showing obvious signs of advanced lymphoma were sacrificed according to human practices. The survival of the test groups was compared with Group 1 or Group 3 and the importance of survival graphs was determined using a logarithmic interval test (Mantel-Cox) using the Prism Graph software.

Animals were treated on day 0 (Figure 4A) or day 0 and day 7 (Figure 4B) with the indicated antibody or the combination of antibodies at the indicated concentration and were evaluated by flow cytometry on day 6 (Figure 4A) or on day 13 (Figure 4B). The exception was that the animals in panel B group "10 mpk CD20 1 mpk 4-1BB s.d." they received the CD20 antibody on days 0 and 7, but 4-1BB only on day 0. The data is represented as a change between animals in the percentage of low B220 cells in relation to the day before enrollment (day - 1) and are calculated using the following formula ((% of low cells in B220 on the indicated study day -% of low cells in B220 the day before enrollment) /% of low cells in B220 on the day before registration) * 100). The statistical significance of the treatment groups compared to the 10 mpk CD20 group was determined using the 1-way ANOVA analysis and the Dunnet multiple comparison test using Graph Pad Prism software; n> 5 / group * p <0.05, *** p <0.005.

Animals were treated with 10 mg / kg of CD20 weekly for 4 weeks, 1 mg / kg of 4-1BB weekly for 2 weeks or a combination of CD20 and the indicated concentration of 4-1BB according to the indicated schedule. Survival days after treatment of the two groups are shown in Figure 5. The average survival was calculated using Graph Pad Prism CD20 = 15 days, 1 mpk 4-1BB = 23.5 days, CD20 0.1 mpk 4- 1BB = 25.5 days, CD20 1 mpk 4-1 BB = 34 days. The statistical importance of the 4-1BB treatment or the combined treatment with CD20 was determined by a logarithmic interval test (Mantel-Cox) using Graph Pad Prism, CD20 vs. software. 4-1BB * p <0.025, CD20 vs. CD20 0.1 mpk 4-1 ^b B p = not significant, CD20 vs. CD20 1 mpk4-1BB *** □□□ p <0.0001.

The results indicate that the treatment of animals with MAB9371 only demonstrated a stabilization of the disease by FACS analysis. However, a significant reduction in circulating tumor was observed for all dose groups combined after treatment (Figure 4).

Raw Sequence Listing

(In amino acid sequences, CDRs are underlined, variable regions in uppercase and constant regions in lowercase)

A. MOR-6032 ANTIBODY

Amino acid sequence of V ^h (SEQ ID NO .: 4):

QVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGGIIP

GFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARKNEEDGGFDHWG

QGTLVTVSS

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 5):

QVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGGIIP

GFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARKNEEDGGFDHWG

QGTLVTVSSastkgpsvfplapcsrstsestaalgcIvkdyfpepvtvswnsgaltsgvhtfpavIqssglysIssvvtvp ssnfgtqtytcnvdhkpsntkvdktverkccvecppcpappvagpsvflfppkpkdtlmisrtpevtcvvvdvshedpevqf nwyvdgvevhnaktkpreeqfnstfrvvsvltvvhqdwlngkeykckvsnkglpapiektisktkgqprepqvytlppsree mtknqvsltclvkgfypsdiavewesngqpennykttppmldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytq kslslspgk

Amino acid sequence of V ^l (SEQ ID NO .: 9):

DIELTQPPSVSVAPGQTARISCSGDNLGDYYASWYQQKPGQAPVLVIYDDSNRP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTWDGTLHFVFGGGTKLTVL

Sequence of full-length light chain amino acids (SEQ ID NO .: 10):

DIELTQPPSVSVAPGQTARISCSGDNLGDYYASWYQQKPGQAPVLVIYDDSNRPSGIPE

RFSGSNSGNTATLTISGTQAEDEADYYCQTWDGTLHFVFGGGTKLTVLqqpkaapsvtlfpp sseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshrsyscqvtheprsgstv

V ^h nucleic acid sequence (SEQ ID NO .: 11):

caggtgcaattggttcagtctggcgcggaagtgaaaaaaccgggcagcagcgtgaaagtgagctgcaaagcct ccggaggcacttttaattcttatgctatttcttgggtgcgccaagcccctgggcagggtctcgagtggatgggcggtatcattcc gggttttggcactgcgaattacgcgcagaagtttcagggccgggtgaccattaccgcggatgaaagcaccagcaccgcgt atatggaactgagcagcctgcgtagcgaagatacggccgtgtattattgcgcgcgtaagaatgaggaggatggtggttttga tcattggggccaaggcaccctggtgacggttagctca

Full length heavy chain nucleic acid sequence (SEQ ID NO .: 13): caggtgcaattggttcagtctggcgcggaagtgaaaaaaccgggcagcagcgtgaaagtgagctgcaaagcct ccggaggcacttttaattcttatgctatttcttgggtgcgccaagcccctgggcagggtctcgagtggatgggcggtatcattcc gggttttggcactgcgaattacgcgcagaagtttcagggccgggtgaccattaccgcggatgaaagcaccagcaccgcgt atatggaactgagcagcctgcgtagcgaagatacggccgtgtattattgcgcgcgtaagaatgaggaggatggtggttttga tcattggggccaaggcaccctggtgacggttagctcagcctccaccaagggcccatcggtcttccccctggcgccctgctcc aggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtgga actcaggcgctctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtagt gaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtgga caagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttcc ccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaaga aacagcacgttccgtgtggtcagcgtcctcaccgtcgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggt ccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttc ctccaacaaaggcctcccagccccca tcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgt acaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccag cgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacacctcccatgctggactccga cggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatg catgaggctctgcacaaccactacacacagaagagcctctccctgtctccgggtaaa

Nucleic acid sequence of V ^l (SEQ ID NO .: 12):

Gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatcttggtgattattatgcttcttggtaccagcagaaacccgggcaggcgccagttcttgtgatttatgatgattctaatcgtccct caggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcggaag acgaagcggattattattgccagacttgggatggtactcttcattttgtgtttggcggcggcacgaagttaaccgttctt

Sequence of full length light chain nucleic acids (SEQ ID NO .: 14):

Gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgataatcttg gtgattattatgcttcttggtaccagcagaaacccgggcaggcgccagttcttgtgatttatgatgattctaatcgtccctcaggc atcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcggaagacgaa gcggattattattgccagacttgggatggtactcttcattttgtgtttggcggcggcacgaagttaaccgttcttggtcagcccaag gctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaaggccacactggtgtgtctcataagtga cttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaaggcgggagtggagaccaccacacc gctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacagaatgttca ctccaaacaaagcaacaacaagtacgcggccagcagctacctgagcctgacgcctgagcagtggaagtcccacagaa

B. MOR 7361 antibody

Amino acid sequence of V ^h (SEQ ID NO .: 18)

QVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMHWVRQAPGKGLEWVSVIS

GSGSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLYAQFEGDFWG

QGTLVTVSS

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 19):

QVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMHWVRQAPGKGLEWVSVIS

GSGSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLYAQFEGDFWG

QGTLVTVSSastkgpsvfplapcsrstsestaalgcIvkdyfpepvtvswnsgaltsgvhtfpavIqssglysIssvvtvp ssnfgtqtytcnvdhkpsntkvdktverkccvecppcpappvagpsvflfppkpkdtlmisrtpevtcvvvdvshedpevqf nwyvdgvevhnaktkpreeqfnstfrvvsvltvvhqdwlngkeykckvsnkglpapiektisktkgqprepqvytlppsree mtknqvsltclvkgfypsdiavewesngqpennykttppmldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytq kslslspgk

Amino acid sequence of V ^l (SEQ ID NO .: 23):

DIELTQPPSVSVAPGQTARISCSGDNIGSKYVSWYQQKPGQAPVLVIYSDSERP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSWDGS-ISRVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO: 24):

DIELTQPPSVSVAPGQTARISCSGDNIGSKYVSWYQQKPGQAPVLVIYSDSERP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCQSWDGSISRVFGGGTKLTVLqqpkaap svtlfppsseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshgysst

V ^h nucleic acid sequence (SEQ ID NO .: 25):

Caggtgcaattggtggaaagcggcggcggcctggtgcaaccgggcggcagcctgcgtctgagctgcgcggcc tccggatttaccttttctgattattatatgcattgggtgcgccaagcccctgggaagggtctcgagtgggtgagcgttatctctggtt aatgaacagcctgcgtgcggaagatacggccgtgtattattgcgcgcgtctttatgctcagtttgagggtgatttttggggccaa ctggtagcaatacctattatgcggatagcgtgaaaggccgttttaccatttcacgtgataattcgaaaaacaccctgtatctgca ggcaccctggtgacggttagctca

Full length heavy chain nucleic acid sequence (SEQ ID NO .: 27):

caggtgcaattggtggaaagcggcggcggcctggtgcaaccgggcggcagcctgcgtctgagctgcgcggcctccggatt taccttttctgattattatatgcattgggtgcgccaagcccctgggaagggtctcgagtgggtgagcgttatctctggttctggtag caatacctattatgcggatagcgtgaaaggccgttttaccatttcacgtgataattcgaaaaacaccctgtatctgcaaatgaa cagcctgcgtgcggaagatacggccgtgtattattgcgcgcgtctttatgctcagtttgagggtgatttttggggccaaggcacc ctggtgacggttagctcagcctccaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagc acagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccag cggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtagtgaccgtgccctccagcaact tcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaat gttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacac cctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactg gtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtc agcgtcctcaccgtcgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccag cccccatcg agaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccggg gagcaatgggcagccggagaacaactacaagaccacacctcccatgctggactccgacggctccttcttcctctacagca aggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtggga acacaca agctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccact

gaagagcctctccctgtctccgggtaaa

Nucleic acid sequence of V ^l (SEQ ID NO .: 26) gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatattggttctaagtatgtttcttggtaccagcagaaacccgggcaggcgccagttcttgtgatttattctgattctgagcgtccct caggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcggaag acgaagcggattattattgccagtcttgggatggttctatttctcgtgtgtttggcggcggcacgaagttaaccgtcctaggtcag

Sequence of full length light chain nucleic acids (SEQ ID NO .: 28)

gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatattggttctaagtatgtttcttggtaccagcagaaacccgggcaggcgccagttcttgtgatttattctgattctgagcgtccct caggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcggaag acgaagcggattattattgccagtcttgggatggttctatttctcgtgtgtttggcggcggcacgaagttaaccgtcctaggtcag

C. MOR 7480 antibody

Amino acid sequence of V ^h (SEQ ID NO .: 32)

QVQLVQSGAEVKKPGESLKISCKGSGYSFSTYWISWVRQMPGKGLEWMGKIYP

GDSYTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGYGIFDYWGQGTL

VTVSS

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO.:33)

QVQLVQSGAEVKKPGESLKISCKGSGYSFSTYWISWVRQMPGKGLEWMGKIYP

GDSYTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGYGIFDYWGQGTL

VTVSSastkgpsvfplapcsrstsestaalgcIvkdyfpepvtvswnsgaltsgvhtfpavIqssglysIssvvtvpssnfgt qtytcnvdhkpsntkvdktverkccvecppcpappvagpsvflfppkpkdtlmisrtpevtcvvvdvshedpevqfnwyvcl gvevhnaktkpreeqfnstfrvvsvltvvhqdwlngkeykckvsnkglpapiektisktkgqprepqvytlppsreemtknq vsltclvkgfypsdiavewesngqpennykttppmldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslsls pgk

Amino acid sequence of V ^l (SEQ ID NO.:37)

DIELTQPPSVSVAPGQTARISCSGDNIGDQYAHWYQQKPGQAPVWIYQDKNRP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCATYTGFGSLAVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO 38):

DIELTQPPSVSVAPGQTARISCSGDNIGDQYAHWYQQKPGQAPVWIYQDKNRPSGIPE

RFSGSNSGNTATLTISGTQAEDEADYYCATYTGFGSLAVFGGGTKLTVLqgpkaapsvtlfpp sseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshrsyscktvthegstyscktvthegstysvt

V ^h nucleic acid sequence (SEQ ID NO.:39)

caggtgcaattggttcagagcggcgcggaagtgaaaaaaccgggcgaaagcctgaaaattagctgcaaaggtt ccggatattccttttctacttattggatttcttgggtgcgccagatgcctgggaagggtctcgagtggatgggcaagatctatccg ggtgatagctataccaattattctccgagctttcagggccaggtgactattagcgcggataaaagcattagcaccgcgtatctt caatggagcagcctgaaagcgagcgatacggccatgtattattgtgcgcgtggttatggtatttttgattattggggccaaggc accctggtcaccgtctcctca

Nucleic acid sequence of heavy chain full length (IgG2) (SEQ ID NO .: 41) caggtgcaattggttcagagcggcgcggaagtgaaaaaaccgggcgaaagcctgaaaattagctgcaaaggttccggat attccttttctacttattggatttcttgggtgcgccagatgcctgggaagggtctcgagtggatgggcaagatctatccgggtgat agctataccaattattctccgagctttcagggccaggtgactattagcgcggataaaagcattagcaccgcgtatcttcaatgg agcagcctgaaagcgagcgatacggccatgtattattgtgcgcgtggttatggtatttttgattattggggccaaggcaccctg gtcaccgtctcctcagcctccaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcaca gcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcgg cgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtagtgaccgtgccctccagcaacttcg gcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgtt gtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccc tcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggt acgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcag cgtcctcaccgtcgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcc cccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccggga ggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggag gctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccacta agcaatgggcagccggagaacaactacaagaccacacctcccatgctggactccgacggctccttcttcctctacagcaa cacacagaagagcctctccctgtctccgggtaaa

V ^l nucleic acid sequence (SEQ ID NO .: 40)

Gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatattggtgatcagtatgctcattggtaccagcagaaacccgggcaggcgccagttgttgtgatttatcaggataagaatcgt ccctcaggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcgg aagacgaagcggattattattgcgctacttatactggttttggttctcttgctgtgtttggcggcggcacgaagttaaccgtccta

Sequence of full length light chain nucleic acids (SEQ ID NO .: 42)

gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatattggtgatcagtatgctcattggtaccagcagaaacccgggcaggcgccagttgttgtgatttatcaggataagaatcgt ccctcaggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcgg aagacgaagcggattattattgcgctacttatactggttttggttctcttgctgtgtttggcggcggcacgaagttaaccgtccta

D. MOR 7480.1 Antibody

Amino acid sequence of V ^h (SEQ ID NO .: 43):

EVQLVQSGAEVKKPGESLRISCKGSGYSFSTYWISWVRQMPGKGLEWMGKIYP

GDSYTNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGYGIFDYWGQGTL

VTVSS

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 44):

EVQLVQSGAEVKKPGESLRISCKGSGYSFSTYWISWVRQMPGKGLEWMGKIYPGDSY

TNYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGYGIFDYWGQGTLVTVS

Sastkgpsvfplapcsrstsestaalgclvkdyfpepvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssnfgtqtytcn vdhkpsntkvdktverkccvecppcpappvagpsvflfppkpkdtlmisrtpevtcvvvdvshedpevqfnwyvdgvevh naktkpreeqfnstfrvvsvltvvhqdwlngkeykckvsnkglpapiektisktkgqprepqvytlppsreemtknqvsltclv kgfypsdiavewesngqpennykttppmldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspgk

Amino acid sequence of V ^l (SEQ ID NO .: 45):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVLVIYQDKNR

PSGIPERFSGSNSGNTATLTISGTQAMDEADYYCATYTGFGSLAVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO .: 46):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVLVIYQDKNR

PSGlPERFSGSNSGNTATLTISGTQAMDEADYYCATYTGFGSLAVFGGGTKLTVLqgDka apsvtlfppsseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwthegrsvecktvpksgrsysc

V ^h nucleic acid sequence (SEQ ID NO.:47):

gaggtgcaattggttcagagcggcgcggaagtgaaaaaaccgggcgaaagcctgaggattagctgcaaaggt tccggatattccttttctacttattggatttcttgggtgcgccagatgcctgggaagggtctcgagtggatgggcaagatctatccg ggtgatagctataccaattattctccgagctttcagggccaggtgactattagcgcggataaaagcattagcaccgcgtatctt caatggagcagcctgaaagcgagcgatacggccatgtattattgtgcgcgtggttatggtatttttgattattggggccaaggc accctggtcaccgtctcctca

Sequence of full-length heavy chain nucleic acids (lgG2) (SEQ ID NO .: 49):

gaggtgcaattggttcagagcggcgcggaagtgaaaaaaccgggcgaaagcctgaggattagctgcaaaggt tccggatattccttttctacttattggatttcttgggtgcgccagatgcctgggaagggtctcgagtggatgggcaagatctatccg

ggtgatagctataccaattattctccgagctttcagggccaggtgactattagcgcggataaaagcattagcaccgcgtatctt caatggagcagcctgaaagcgagcgatacggccatgtattattgtgcgcgtggttatggtatttttgattattggggccaaggc accctggtcaccgtctcctcagcctccaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgaga gcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgacc agcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctcagcagcgtagtgaccgtgccctccagca aatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaagga acttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgca caccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttca actggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgt ggtcagcgtcctcaccgtcgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctc ccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcc cgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagt gggagagcaatgggcagccgg agaacaactacaagaccacacctcccatgctggactccgacggctccttcttcctctac agcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaa ccactacacacagaagagctctg

V nucleic acid sequence ^l (SEQ ID NO.:48):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggc gacaacatcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtgctggtgatctaccaggac aagaaccggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcg gcacccaggccatggacgaggccgactactactgcgccacctacaccggcttcggcagcctggccgtgttcggcggagg gaccaagctgaccgtccta

Sequence of full-length light chain nucleic acids (SEQ ID NO .: 50):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggcgacaac atcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtgctggtgatctaccaggacaagaac cggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcggcaccca ggccatggacgaggccgactactactgcgccacctacaccggcttcggcagcctggccgtgttcggcggagggaccaag ctgaccgtcctaggtcagcccaaggctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaagg ccacactggtgtgtctcataagtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaagg c99ga9t99a9accaccacaccctccaaacaaa9caacaacaagtac9c99cca9ca9ctacct9agcct9ac9cct gagcagtggaagtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccc tacagaatgttca

E. MOR 7480.2 Antibody

Amino acid sequence of V ^h (SEQ ID NO .: 43): (same as MOR-7480.1)

Full-length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 44): (same as IgG2 MOR-7480.1)

Amino acid sequence of V ^l (SEQ ID NO .: 51):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVWIYQDKNR

PSGIPERFSGSNSGNTATLTISGTQAMDEADYYCATYTGFGSLAVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO 52):

SYE LTQ P PSVSVSPGQTASIT CSGDNIGDQYAH WY QQ KPGQS PVWIYQDKNRPSGIP

ERFSGSNSGNTATLTISGTQAMDEADYYCATYTGFGSLAVFGGGTKLTVLgqpkaapsvtlf ppsseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshrsyscva

V ^h nucleic acid sequence (SEQ ID NO.:47): (same as MOR-7480.1)

Full length heavy chain nucleic acid sequence (IgG2) (SEQ ID NO .: 49): (same as IgG2 MOR-7480.1)

Nucleic acid sequence of V ^l (SEQ ID NO .: 53):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggc gacaacatcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtggtggtgatctaccaggac aagaaccggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcg gcacccaggccatggacgaggccgactactactgcgccacctacaccggcttcggcagcctggccgtgttcggcggagg gaccaagctgaccgtccta

Sequence of full length light chain nucleic acids (SEQ ID NO .: 54):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggcgacaac atcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtggtggtgatctaccaggacaagaac cggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcggcaccca ggccatggacgaggccgactactactgcgccacctacaccggcttcggcagcctggccgtgttcggcggagggaccaag ctgaccgtcctaggtcagcccaaggctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaagg

ccacactggtgtgtctcataagtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaagg cgggagtggagaccaccacaccctccaaacaaagcaacaacaagtacgcggccagcagctacctgagcctgacgcct gagcagtggaagtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccc tacagaatgttca

F. MOR-7483 Antibody

Amino acid sequence of V ^h (SEQ ID NO .: 32): (Same as MOR 7480)

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 33): (Same as IgG2 MOR 7480)

Amino acid sequence of V ^l (SEQ ID NO .: 56):

DIELTQPPSVSVAPGQTARISCSGDNIGDQYAHWYQQKPGQAPVVVIYQDKNRP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTYTFVGFTTVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO: 57)

DIELTQPPSVSVAPGQTARISCSGDNIGDQYAHWYQQKPGQAPVWIYQDKNRP

SGIPERFSGSNSGNTATLTISGTQAEDEADYYCSTYTFVGFTTVFGGGTKLTVLqgpkaap svtlfppsseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshgysstl

V ^h nucleic acid sequence (SEQ ID NO .: 39): (Same as MOR 7480)

Sequence of full-length heavy chain nucleic acids (IgG2) (SEQ ID NO .: 41): (Same as IgG2 MOR 7480)

Nucleic acid sequence of V ^l (SEQ ID NO .: 58):

Gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgat aatattggtgatcagtatgctcattggtaccagcagaaacccgggcaggcgccagttgttgtgatttatcaggataagaatcgt ccctcaggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcgg aagacgaagcggattattattgctctacttatacttttgttggttttactactgtgtttggcggcggcacgaagttaaccgtccta

Sequence of full-length light chain nucleic acids (SEQ ID NO .: 59):

Gatatcgaactgacccagccgccttcagtgagcgttgcaccaggtcagaccgcgcgtatctcgtgtagcggcgataatattg gtgatcagtatgctcattggtaccagcagaaacccgggcaggcgccagttgttgtgatttatcaggataagaatcgtccctca ggcatcccggaacgctttagcggatccaacagcggcaacaccgcgaccctgaccattagcggcactcaggcggaagac gaagcggattattattgctctacttatacttttgttggttttactactgtgtttggcggcggcacgaagttaaccgtcctaggtcagcc caaggctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaaggccacactggtgtgtctcataa gtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaaggcgggagtggagaccacca caccctccaaacaaagcaacaacaagtacgcggccagcagctacctgagcctgacgcctgagcagtggaagtcccaca gaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacagaatgttca

G. MOR-7483.1 Antibody

V amino acid sequence ^h (SEQ ID NO .: 43): (same as MOR 7480.1)

Full-length heavy chain amino acid sequence (IgG2) (SEQ ID NO .: 44): (same as IgG2 MOR-7480.1)

Amino acid sequence of V ^l (SEQ ID NO .: 60):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVLVIYQDKNR

PSGIPERFSGSNSGNTATLTISGTQAMDEADYYCSTYTFVGFTTVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO .: 61):

DIELTQPPSVSVAPGQTARISCSGDNIGDQYAHWYQQKPGQAPVWIYQDKNRPSGIPE

RFSGSNSGNTATLTISGTQAEDEADYYCSTYTFVGFTTVFGGGTKLTVLagpkaapsvtlfpp

sseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshrsyscqvthegstv

ektvaptecs

V ^h nucleic acid sequence (SEQ ID NO .: 47): (same as MOR-7480.1)

Full length heavy chain nucleic acid sequence (IgG2) (SEQ ID NO .: 49): (same as IgG2 MOR-7480.1)

Nucleic acid sequence of V ^l (SEQ ID NO .: 62):

Agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggc

gacaacatcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtgctggtgatctaccaggac

aagaaccggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcg

gcacccaggccatggacgaggccgactactactgctctacttatacttttgttggttttactactgtgttcggcggagggaccaa

gctgaccgtccta

Sequence of full-length light chain nucleic acids (SEQ ID NO .: 63):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggcgacaac

atcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtgctggtgatctaccaggacaagaac

cggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcggcaccca

ggccatggacgaggccgactactactgctctacttatacttttgttggttttactactgtgttcggcggagggaccaagctgacc

gtcctaggtcagcccaaggctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaaggccacac

tggtgtgtctcataagtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaaggcggga

gtggagaccaccacaccctccaaacaaagcaacaacaagtacgcggccagcagctacctgagcctgacgcctgagca

gtggaagtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacag

aatgttca

H. MOR-7483.2 Antibody

Amino acid sequence of V ^h (SEQ ID NO .: 43): (same as MOR-7480.1)

Full length heavy chain amino acid sequence (IgG2) (SEQ ID NO.:44): (same as IgG2 MOR-7480.1)

Amino acid sequence of V ^l (SEQ ID NO .: 64):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVWIYQDKNR

PSGIPERFSGSNSGNTATLTISGTQAMDEADYYCSTYTFVGFTTVFGGGTKLTVL

Full length light chain amino acid sequence (SEQ ID NO: 65):

SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVWIYQDKNRPSGIP

ERFSGSNSGNTATLTISGTQAMDEADYYCSTYTFVGFTTVFGGGTKLTVLqgDkaaDSvtlfp psseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltpeqwkshrsyscqvtegst vektcva

V ^h nucleic acid sequence (SEQ ID NO .: 47): (same as MOR-7480.1)

Full length heavy chain nucleic acid sequence (IgG2) (SEQ ID NO .: 49): (same as IgG2 MOR-7480.1)

Nucleic acid sequence of V ^l (SEQ ID NO .: 66):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggc gacaacatcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtggtggtgatctaccaggac aagaaccggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcg gcacccaggccatggacgaggccgactactactgctctacttatacttttgttggttttactactgtgttcggcggagggaccaa gctgaccgtccta

Sequence of full length light chain nucleic acids (SEQ ID NO .: 67):

agctacgagctgacccagccccccagcgtgtccgtgagccctggccagaccgccagcatcacctgcagcggcgacaac atcggcgaccagtacgcccactggtatcagcagaagcccggccagagccccgtggtggtgatctaccaggacaagaac cggcccagcggcatccccgagcggttcagcggcagcaacagcggcaacaccgccaccctgaccatcagcggcaccca ggccatggacgaggccgactactactgctctacttatacttttgttggttttactactgtgttcggcggagggaccaagctgacc gtcctaggtcagcccaaggctgccccctcggtcactctgttcccaccctcctctgaggagcttcaagccaacaaggccacac tggtgtgtctcataagtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaaggcggga gtggagaccaccacaccctccaaacaaagcaacaacaagtacgcggccagcagctacctgagcctgacgcctgagca gtggaagtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacag aatgttca

I. Amino acid sequence of human 4-1BB (SEQ ID NO: 68):

mgnscynivatlllvlnfertrslqdpcsncpagtfcdnnrnqicspcppnsfssaggqrtcdicrqckgvfrtrkecs stsnaecdctpgfhclgagcsmceqdckqgqeltkkgckdccfgtfndqkrgicrpwtncsldgksvlvngtkerdvvcgps padlspgassvtppaparepghspqiisfflaltstallfllffltlrfsvvkrgrkkllyifkqpfnnrpvqttqeedgcscrfpeeeegg cel

J. Amino acid sequence of the human IgG1 constant region (SEQ ID NO: 69):

astkgpsvfplapsskstsggtaalgclvkdyfpepvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtq tyicnvnhkpsntkvdkkvepkscdkthtcppcpapellggpsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwy vdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpapiektiskakgqprepqvytlppsreemtk nqvsltclvkgfypsdiavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslsl spgk

K. Nucleic acid sequence of the human IgG1 constant region (SEQ ID NO: 70):

GCCT CCACCAAGGGCCCAT CG GT CTT CCCCCTGGCACCCT CCT CCAAGAGCACCT

CTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGT

GACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCT

GT CCTACAGT CCT CAGGACT CTACT CCCT CAGCAGCGTAGT GACCGT GCCCT CCAG

CAGCTT G GGCACCCAG ACCTACAT CT GCAACGT GAAT CACAAGCCCAGCAACACCA

AGGTGGACAAGAAAGTT GAGCCCAAAT CTT GT GACAAAACT CACACAT GCCCACCG

T GCCCAGCACCT GAACT CCT GGGGGGACCGT CAGT CTT CCT CTT CCCCCCAAAACC

CAAGGACACCCT CAT GAT CT CCCGGACCCCT GAGGT CACAT GCGTGGT G GTGGAC

GT GAGCCACGAAGACCCT GAGGT CAAGTT CAACT GGTACGT GGACGGCGT GGAGG

TGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGT

GGT CAGCGT CCT CACCGT CCT GCACCAGGACTGGCT GAAT GGCAAGGAGTACAAG

T GCAAG GT CT CCAACAAAGCCCT CCCAGCCCCCAT CG AGAAAACCAT CTCCAAAGC

CAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAG

AT GACCAAGAACCAGGT CAGCCT GACCT GCCTGGT CAAAGGCTT CTAT CCCAGCGA

CATCGCCGT G GAGT GG GAG AGCAAT G GGCAGCCG GAG AACAACTACAAGACCACG

CCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGA

CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCT

CT GCACAACCACTACACGCAG AAGAGCCT CTCCCTGTCT CCG GGTAAA

L. Amino acid sequence of the human IgG2 constant region (SEQ ID NO: 71)

astkgpsvfplapcsrstsestaalgclvkdyfpepvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssnfgtq tytcnvdhkpsntkvdktverkccvecppcpappvagpsvflfppkpkdtlmisrtpevtcvvvdvshedpevqfnwyvdg vevhnaktkpreeqfnstfrvvsvltvvhqdwlngkeykckvsnkglpapiektisktkgqprepqvytlppsreemtknqvs k Itclvkgfypsdiavewesngqpennykttppmldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspg

M. Nucleic acid sequence of the human IgG2 constant region (SEQ ID NO: 72):

GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCT

CCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGT

GACGGT GT CGT GGAACT CAGGCGCT CT GACCAGCGGCGT GCACACCTT CCCGGCT

GT CCTACAGT CCT CAGGACT CTACT CCCT CAGCAGCGTAGT GACCGTGCCCT CCAG

CAACTTCGGCACCCAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCA

AGGTGGACAAGACAGTT GAGCGCAAAT GTT GT GTCGAGT GCCCACCGTGCCCAGC

ACCACCT GT G GCAGG ACCGT CAGT CTT CCT CTT CCCCCCAAAACCCAAG GACACCC

T CAT GAT CT CCCGGACCCCT GAGGT CACGT GCGTGGT GGT GGACGT GAGCCACGA

AGACCCCGAGGT CCAGTT CAACTGGTACGT GGACGGCGT GGAGGT GCATAAT GCC

AAGACAAAGCCACGGGAGGAGCAGTT CAACAGCACGTT CCGT GT GGT CAGCGT CC

TCACCGTCGTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTC

CAACAAAGGCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGGCAGC

CCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAA

CCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTG

GAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACACCTCCCATGC

T G GACT CCG ACG GCT CCTT CTT CCT CTACAGCAAGCT CACCGT GG ACAAG AGCAGG

T G GCAGCAGG GG AACGT CTT CT CAT GCTCCGT GAT GCAT GAG GCT CT GCACAACCA

CTACACACAGAAG AGCCT CTCCCTGTCTCCGG GTAAA

N. Amino acid sequence of the human lambda light chain constant region (SEQ ID NO: 73):

gqpkaapsvtlfppsseelqankatlvclisdfypgavtvawkadsspvkagvetttpskqsnnkyaassylsltp eqwkshrsyscqvthegstvektvaptecs

O. Nucleic acid sequence of the human lambda light chain constant region (SEQ ID NO: 74):

GGT CAGCCCAAGGCT GCCCCCTCGGT CACTCT GTT CCCACCCT CCT CT GAGGAGC

TT CAAGCCAACAAG GCCACACT G GTGTGTCT CATAAGT G ACTT CT ACCCGG GAGCC

GTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACC

ACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTACCTGAGCCT

GACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAA

GGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA

P. ANTIBODY P-Cadherina (g-194-g09 or anti-P-cadherina1)

Amino acid sequence of H-CDR1 (SEQ ID NO: 75):

SYAMS

Amino acid sequence of H-CDR2 (SEQ ID NO: 76):

AISGSGGSTYYADSVKG

Amino acid sequence of H-CDR3 (SEQ ID NO: 77):

TNSAKFDP

Amino acid sequence of L-CDR1 (SEQ ID NO: 78):

TGTSNDVGAYNYVS

Amino acid sequence of L-CDR2 (SEQ ID NO: 79):

EVNKRPS

Amino acid sequence of L-CDR3 (SEQ ID NO: 80):

SSYTMGSTFM L

Claims (6)

1. An anti-4-1BB antibody, or an antigen binding portion thereof, for use in the treatment of cancer in combination with an anti-CD20 antibody, or an antigen binding portion thereof, wherein said antibody anti-4-1BB, or the antigen binding portion thereof, comprises: (a) an H-CDR1 as set forth in SEQ ID NO: 29; (b) an H-CDR2 as set forth in SEQ ID NO: 30; (c) an H-CDR3 as set forth in SEQ ID NO: 31; (d) an L-CDR1 as set forth in SEQ ID NO: 34; (e) an L-CDR2 as set forth in SEQ ID NO: 35; Y (f) an L-CDR3 as set forth in SEQ ID NO: 36, and wherein the anti-CD20 antibody, or antigen binding portion thereof, is administered in a dose of 3-15 mg / kg. 2. The anti-4-1BB antibody, or antigen binding portion thereof, for the use of claim 1, wherein said anti-CD20 antibody, or antigen binding portion thereof, comprises the 6 CDRs of rituximab. 3. The anti-4-1BB antibody, or antigen binding portion thereof, for the use of claim 1 or 2, wherein said anti-4-1BB antibody, or antigen binding portion thereof, comprises a Vh region comprising the amino acid sequence set forth in SEQ ID NO: 43 and a Vl region comprising the amino acid sequence set forth in SEQ ID NO: 45. 4. The anti-4-1BB antibody, or antigen binding portion thereof, for the use of any one of claims 1 to 3, wherein said anti-CD20 antibody, or antigen binding portion thereof, comprises the V ^h region of rituximab and the V ^l region of rituximab. 5. The anti-4-1BB antibody, or antigen binding portion thereof, for the use of any of claims 1 to 4, wherein said anti-4-1BB antibody, or antigen binding portion thereof , comprises an amino acid sequence of the heavy chain as set forth in SEQ ID NO: 44 and also comprises an amino acid sequence of the light chain set forth in SEQ ID NO: 46, with the proviso that the lysine residue of the C-terminal end of SEQ ID NO: 44 is optionally absent. 6. The anti-4-1BB antibody, or antigen binding portion thereof, for the use of any one of claims 1 to 5, wherein said anti-CD20 antibody, or antigen binding portion thereof, comprises the amino acid sequence of the rituximab heavy chain and the amino acid sequence of the rituximab light chain.